Antitumoral Compounds

ABSTRACT

A compound of general formula I, wherein X, R 1 -R 4  take various meanings, for use in the treatment of cancer.

FIELD OF THE INVENTION

The present invention relates to synthetic analogues of theecteinascidins, particularly of ecteinascidin 736 (ET-736),pharmaceutical compositions containing them, methods for theirmanufacture and their use as antitumoral agents.

BACKGROUND OF THE INVENTION

The ecteinascidins are exceedingly potent antitumor agents isolated fromthe marine tunicate Ecteinascidia turbinata. One of these compounds,ET-743 of formula:

is being employed as an anticancer medicament, under the internationalnonproprietary name (INN) trabectedin, for the treatment of patientswith advanced and metastatic soft tissue sarcoma (STS) after failure ofanthracyclines and ifosfamide, or who are unsuited to receive suchagents, and for the treatment of relapsed platinum-sensitive ovariancancer in combination with pegylated liposomal doxorubicin.

Ecteinascidin 736 (ET-736) was first discovered by Rinehart and featuresa tetrahydro-β-carboline unit in place of the tetrahydroisoquinolineunit more usually found in the ecteinascidin compounds isolated fromnatural sources; See for example Sakai et al., Proc. Natl. Acad. Sci.USA 1992, vol. 89, 11456-11460.

U.S. Pat. No. 5,149,804 describes Ecteinascidin 736 (ET-736), isolatedfrom the Caribbean tunicate Ecteinascidia turbinata, and it structure.ET-736 protects mice in vivo at very low concentrations against P388lymphoma, B16 melanoma, and Lewis lung carcinoma.

WO03014127 describes several synthetic analogues of ET-736 and theircytotoxic activity against tumoral cells. In particular, WO03014127describes compounds A to D together with their cytotoxic activityagainst a panel of cancer cell lines.

Another compound described in this patent application, PM01183, iscurrently in clinical trials for the treatment of cancer. PM01183 hasthe following chemical structure:

PM01183 has demonstrated a highly potent in vitro activity against solidand non-solid tumour cell lines as well as a significant in vivoactivity in several xenografted human tumor cell lines in mice, such asthose for breast, kidney and ovarian cancer. PM01183 exerts itsanticancer effects through the covalent modification of guanines in theDNA minor groove that eventually give rise to DNA double-strand break,S-phase arrest and apoptosis in cancer cells.

Despite the positive results obtained in clinical applications inchemotherapy, the search in the field of ecteinascidin compounds isstill open to the identification of new compounds with optimal featuresof activity, selectivity toward the tumour, with a reduced systemictoxicity and/or improved pharmacokinetic properties.

SUMMARY OF THE INVENTION

In a first aspect of the present invention there is provided a compoundof formula I or a pharmaceutically acceptable salt or ester thereof:

wherein:

X is —NH— or —O—; R₁ is —OH or —CN;

R₂ is a —C(═O)R^(a) group;R₃ is hydrogen or a —OR^(b) group;R₄ is selected from hydrogen, —CH₂OH, —CH₂OC(═O)R^(c), —CH₂NH₂, and—CH₂NHProt^(NH);R^(a) is selected from hydrogen, substituted or unsubstituted C₁-C₁₂alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl, and substituted orunsubstituted C₂-C₁₂ alkynyl;R^(b) is selected from substituted or unsubstituted C₁-C₁₂ alkyl,substituted or unsubstituted C₂-C₁₂ alkenyl, and substituted orunsubstituted C₂-C₁₂ alkynyl;R^(c) is selected from substituted or unsubstituted C₁-C₁₂ alkyl,substituted or unsubstituted C₂-C₁₂ alkenyl, and substituted orunsubstituted C₂-C₁₂ alkynyl; andProt^(NH) is a protecting group for amino,with the proviso that when R₄ is hydrogen then X is —O—.

There is also provided a compound of formula IC, or a pharmaceuticallyacceptable salt or ester thereof:

wherein:

-   -   X is —NH—;    -   R₁ is —OH or —CN;    -   R₂ is a —C(═O)R^(a) group;    -   R₃ is hydrogen or a —OR^(b) group;    -   R₄ is selected from —CH₂OH, —CH₂O—(C═O)R^(c), —CH₂NH₂ and        —CH₂NHProt^(NH);    -   R^(a) is selected from hydrogen, substituted or unsubstituted        C₁-C₁₂ alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl, and        substituted or unsubstituted C₂-C₁₂ alkynyl;    -   R^(b) is selected from substituted or unsubstituted C₁-C₁₂        alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl and        substituted or unsubstituted C₂-C₁₂ alkynyl;    -   R^(c) is selected from substituted or unsubstituted C₁-C₁₂        alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl, and        substituted or unsubstituted C₂-C₁₂ alkynyl; and    -   Prot^(NH) is a protecting group for amino.

There is also provided a compound of formula ID, or a pharmaceuticallyacceptable salt or ester thereof:

wherein:

-   -   X is —O—;    -   R₁ is —OH or —CN;    -   R₂ is a —C(═O)R^(a) group;    -   R₃ is hydrogen or a —OR^(b) group;    -   R₄ is selected from hydrogen, —CH₂OH, —CH₂O—(C═O)R^(c), —CH₂NH₂        and —CH₂NHProt^(NH);    -   R^(a) is selected from hydrogen, substituted or unsubstituted        C₁-C₁₂ alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl, and        substituted or unsubstituted C₂-C₁₂ alkynyl;    -   R^(b) is selected from substituted or unsubstituted C₁-C₁₂        alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl and        substituted or unsubstituted C₂-C₁₂ alkynyl;    -   R^(c) is selected from substituted or unsubstituted C₁-C₁₂        alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl, and        substituted or unsubstituted C₂-C₁₂ alkynyl; and    -   Prot^(NH) is a protecting group for amino.

There is also provided a compound of formula IE, or a pharmaceuticallyacceptable salt or ester thereof:

wherein:

-   -   X is —NH— or —O—;    -   R₁ is —OH or —CN;    -   R₂ is a —C(═O)R^(a) group;    -   R₃ is hydrogen or a —OR^(b) group;    -   R₄ is selected from —CH₂NH₂ and —CH₂NHProt^(NH);    -   R^(a) is selected from hydrogen, substituted or unsubstituted        C₁-C₁₂ alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl, and        substituted or unsubstituted C₂-C₁₂ alkynyl;    -   R^(b) is selected from substituted or unsubstituted C₁-C₁₂        alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl and        substituted or unsubstituted C₂-C₁₂ alkynyl; and    -   Prot^(NH) is a protecting group for amino.

There is also provided a compound of formula IA or a pharmaceuticallyacceptable salt or ester thereof:

wherein:

-   -   X is —NH— or —O—;    -   R₁ is —OH or —CN;    -   R₂ is a —C(═O)R^(a) group;    -   R₃ is hydrogen;    -   R₄ is selected from hydrogen, —CH₂OH, —CH₂O—(C═O)R^(c), —CH₂NH₂        and —CH₂NHProt^(NH);    -   R^(a) is selected from hydrogen, substituted or unsubstituted        C₁-C₁₂ alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl, and        substituted or unsubstituted C₂-C₁₂ alkynyl;    -   R^(c) is selected from substituted or unsubstituted C₁-C₁₂        alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl, and        substituted or unsubstituted C₂-C₁₂ alkynyl; and    -   Prot^(NH) is a protecting group for amino;    -   with the proviso that when R₄ is hydrogen then X is —O—.

There is also provided a compound of formula IB or a pharmaceuticallyacceptable salt or ester thereof:

wherein:

-   -   X is —NH— or —O—;    -   R₁ is —OH or —CN;    -   R₂ is a —C(═O)R^(a) group;    -   R₃ is a —OR^(b) group;    -   R₄ is selected from hydrogen, —CH₂OH, —CH₂O—(C═O)R^(c), —CH₂NH₂        and —CH₂NHProt^(NH);    -   R^(a) is selected from hydrogen, substituted or unsubstituted        C₁-C₁₂ alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl, and        substituted or unsubstituted C₂-C₁₂ alkynyl;    -   R^(b) is selected from substituted or unsubstituted C₁-C₁₂        alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl and        substituted or unsubstituted C₂-C₁₂ alkynyl;    -   R^(c) is selected from substituted or unsubstituted C₁-C₁₂        alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl, and        substituted or unsubstituted C₂-C₁₂ alkynyl; and    -   Prot^(NH) is a protecting group for amino;    -   with the proviso that when R₄ is hydrogen then X is —O—.

There is also provided a compound of formula IF or a pharmaceuticallyacceptable salt or ester thereof:

wherein:

-   -   X is —NH— or —O—;    -   R₁ is —OH;    -   R₂ is a —C(═O)R^(a) group;    -   R₃ is hydrogen or a —OR^(b) group;    -   R₄ is selected from hydrogen, —CH₂OH, —CH₂OC(═O)R^(c), —CH₂NH₂,        and —CH₂NHProt^(NH);    -   R^(a) is selected from hydrogen, substituted or unsubstituted        C₁-C₁₂ alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl, and        substituted or unsubstituted C₂-C₁₂ alkynyl;    -   R^(b) is selected from substituted or unsubstituted C₁-C₁₂        alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl, and        substituted or unsubstituted C₂-C₁₂ alkynyl;    -   R^(c) is selected from substituted or unsubstituted C₁-C₁₂        alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl, and        substituted or unsubstituted C₂-C₁₂ alkynyl; and    -   Prot^(NH) is a protecting group for amino,    -   with the proviso that when R₄ is hydrogen then X is —O—.

There is also provided a compound of formula IG or a pharmaceuticallyacceptable salt or ester thereof:

wherein:

X is —NH— or —O—; R₁ is —OH or —CN;

R₂ is acetyl;R₃ is hydrogen or a —OR^(b) group;R₄ is selected from hydrogen, —CH₂OH, —CH₂OC(═O)R^(c), —CH₂NH₂, and—CH₂NHProt^(NH);R^(b) is selected from substituted or unsubstituted C₁-C₁₂ alkyl,substituted or unsubstituted C₂-C₁₂ alkenyl, and substituted orunsubstituted C₂-C₁₂ alkynyl;R^(c) is selected from substituted or unsubstituted C₁-C₁₂ alkyl,substituted or unsubstituted C₂-C₁₂ alkenyl, and substituted orunsubstituted C₂-C₁₂ alkynyl; andProt^(NH) is a protecting group for amino,with the proviso that when R₄ is hydrogen then X is —O—.

In a further aspect of the present invention, there is provided apharmaceutical composition comprising a compound according to thepresent invention and a pharmaceutically acceptable carrier.

In a yet further aspect of the present invention, there is provided adosage form comprising a pharmaceutical composition according to thepresent invention.

In a yet further aspect of the present invention, there is provided acompound, pharmaceutical composition or dosage form according to thepresent invention for use as a medicament.

In a yet further aspect of the present invention, there is provided acompound, pharmaceutical composition or dosage form according to thepresent invention for use in the treatment of cancer.

In a yet further aspect of the present invention, there is provided theuse of a compound, pharmaceutical composition or dosage form accordingto the present invention for the manufacture of a medicament for thetreatment of cancer.

In a yet further aspect of the present invention, there is provided amethod for the prevention or treatment of cancer, comprisingadministering an effective amount of a compound according to the presentinvention, administering an effective amount of a pharmaceuticalcomposition according to the present invention, or administering aneffective amount of a dosage form according to the present invention toa patient in need thereof, notably a human.

In a yet further aspect of the present invention, there is provided theuse of a compound according to the present invention for the treatmentof cancer, or in the preparation of a medicament preferably for thetreatment of cancer.

In a yet further aspect of the present invention, there is provided akit comprising a therapeutically effective amount of a compoundaccording to the present invention and a pharmaceutically acceptablecarrier. The kit is for use in the treatment of cancer.

In a yet further aspect of the present invention, there is provided aprocess for obtaining compounds of formula I or a pharmaceuticallyacceptable salt or ester thereof, compounds of formula IA or apharmaceutically acceptable salt or ester thereof, compounds of formulaIB or a pharmaceutically acceptable salt or ester thereof, compounds offormula IC or a pharmaceutically acceptable salt or ester thereof,compounds of formula ID or a pharmaceutically acceptable salt or esterthereof, compounds of formula IE or a pharmaceutically acceptable saltor ester thereof, compounds of formula IF or a pharmaceuticallyacceptable salt or ester thereof, compounds of formula IG or apharmaceutically acceptable salt or ester thereof; comprising the stepof reacting a compound of formula II with a compound of formula III togive a compound of formula IV:

-   -   wherein (insofar as allowed by possible substituent groups):    -   X is —NH— or —O—;    -   R₂ is a —C(═O)R^(a) group;    -   R₃ is hydrogen or a —OR^(b) group;    -   R₄ is selected from hydrogen, —CH₂OH, —CH₂OC(═O)R^(c), and        —CH₂NHProt^(NH);    -   R^(a) is selected from hydrogen, substituted or unsubstituted        C₁-C₁₂ alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl, and        substituted or unsubstituted C₂-C₁₂ alkynyl;    -   R^(b) is selected from substituted or unsubstituted C₁-C₁₂        alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl, and        substituted or unsubstituted C₂-C₁₂ alkynyl;    -   R^(c) is selected from substituted or unsubstituted C₁-C₁₂        alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl, and        substituted or unsubstituted C₂-C₁₂ alkynyl; and    -   Prot^(NH) is a protecting group for amino;    -   with the proviso that when R₄ is hydrogen then X is —O—.    -   The process may include the further step of replacing the cyano        group in the compound of formula IV with a hydroxy group to give        a compound of formula I, IA, IB, IC, ID, IE, IF, or IG where R₁        is OH.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1. Tumor total diameter evaluation of HT1080 tumors in mice treatedwith placebo, compound C, 4-S, and 12-S.

FIG. 2. Tumor volume evaluation of MDA-MB-231 tumors in mice treatedwith placebo, compound C, 4-S, and 12-S.

FIG. 3. Tumor volume evaluation of H460 tumors in mice treated withplacebo, compound C, 4-S, and 12-S.

FIG. 4. Tumor volume evaluation of H526 tumors in mice treated withplacebo, compound C, 4-S, and 12-S.

FIG. 5. Tumor volume evaluation of H82 tumors in mice treated withplacebo, compound C, 4-S, and 12-S.

FIG. 6. Tumor volume evaluation of A2780 tumors in mice treated withplacebo, compound C, 4-S, and 12-S.

FIG. 7. Tumor volume evaluation of HGC-27 tumors in mice treated withplacebo, compound C, 4-S, and 12-S.

FIG. 8. Tumor total diameter evaluation of HT1080 tumors in mice treatedwith placebo, PM01183 and 4-R.

FIG. 9. Tumor total diameter evaluation of HT1080 tumors in mice treatedwith placebo, PM01183 and Compound D.

FIG. 10. Tumor volume evaluation of MDA-MB-231 tumors in mice treatedwith placebo, PM01183 and 4-R.

FIG. 11. Tumor volume evaluation of MDA-MB-231 tumors in mice treatedwith placebo, PM01183 and Compound D.

FIG. 12. Tumor volume evaluation of H460 tumors in mice treated withplacebo, PM01183 and 4-R.

FIG. 13. Tumor volume evaluation of H460 tumors in mice treated withplacebo, PM01183 and Compound D.

FIG. 14. Tumor volume evaluation of A2780 tumors in mice treated withplacebo, PM01183 and 4-R.

FIG. 15. Tumor volume evaluation of A2780 tumors in mice treated withplacebo, PM01183 and Compound D.

FIG. 16. Tumor volume evaluation of HGC-27 tumors in mice treated withplacebo, PM01183 and 4-R.

FIG. 17. Tumor volume evaluation of HGC-27 tumors in mice treated withplacebo, PM01183 and Compound D.

FIG. 18. Tumor total diameter evaluation of HT1080 tumors in micetreated with placebo, compound D and 12-R.

FIG. 19. Tumor volume evaluation of MDA-MB-231 tumors in mice treatedwith placebo, compound D and 12-R.

FIG. 20. Tumor volume evaluation of H460 tumors in mice treated withplacebo, compound D and 12-R.

FIG. 21. Tumor volume evaluation of H526 tumors in mice treated withplacebo, compound D and 12-R.

FIG. 22. Tumor volume evaluation of H82 tumors in mice treated withplacebo, compound D and 12-R.

FIG. 23. Tumor volume evaluation of A2780 tumors in mice treated withplacebo, compound D and 12-R.

FIG. 24. Tumor volume evaluation of HGC-27 tumors in mice treated withplacebo, compound D and 12-R.

FIG. 25. Tumor total diameter evaluation of HT1080 tumors in micetreated with placebo, PM01183 and 19-S.

FIG. 26. Tumor volume evaluation of MDA-MB-231 tumors in mice treatedwith placebo, PM01183 and 19-S.

FIG. 27. Tumor volume evaluation of H460 tumors in mice treated withplacebo, PM01183 and 19-S.

FIG. 28. Tumor volume evaluation of A2780 tumors in mice treated withplacebo, PM01183 and 19-S.

FIG. 29. Tumor volume evaluation of HGC27 tumors in mice treated withplacebo, PM01183 and 19-S.

FIG. 30. Tumor total diameter evaluation of HT1080 tumors in micetreated with placebo, PM01183 and 19-R.

FIG. 31. Tumor volume evaluation of MDA-MB-231 tumors in mice treatedwith placebo, PM01183 and 19-R.

FIG. 32. Tumor volume evaluation of H460 tumors in mice treated withplacebo, PM01183 and 19-R.

FIG. 33. Tumor volume evaluation of A2780 tumors in mice treated withplacebo, PM01183 and 19-R.

FIG. 34. Tumor volume evaluation of HGC-27 tumors in mice treated withplacebo, PM01183 and 19-R.

FIG. 35. Tumor total diameter evaluation of HT1080 tumors in micetreated with placebo, compound C and 39-S.

FIG. 36. Tumor volume evaluation of MDA-MB-231 tumors in mice treatedwith placebo, compound C and 39-S.

FIG. 37. Tumor volume evaluation of H460 tumors in mice treated withplacebo, compound C and 39-S.

FIG. 38. Tumor volume evaluation of H526 tumors in mice treated withplacebo, compound C and 39-S.

FIG. 39. Tumor volume evaluation of H82 tumors in mice treated withplacebo, compound C and 39-S.

FIG. 40. Tumor volume evaluation of A2780 tumors in mice treated withplacebo, compound C and 39-S.

FIG. 41. Tumor volume evaluation of HGC27 tumors in mice treated withplacebo, compound C and 39-S.

FIG. 42. Tumor total diameter evaluation of HT1080 tumors in micetreated with placebo, compound D and 47-R.

FIG. 43. Tumor volume evaluation of MDA-MB-231 tumors in mice treatedwith placebo, compound D and 47-R.

FIG. 44. Tumor volume evaluation of H460 tumors in mice treated withplacebo, compound D and 47-R.

FIG. 45. Tumor volume evaluation of H526 tumors in mice treated withplacebo, compound D and 47-R.

FIG. 46. Tumor volume evaluation of H82 tumors in mice treated withplacebo, compound D and 47-R.

FIG. 47. Tumor volume evaluation of A2780 tumors in mice treated withplacebo, compound D and 47-R.

FIG. 48. Tumor volume evaluation of HGC27 tumors in mice treated withplacebo, compound D and 47-R.

FIG. 49. Tumor total diameter evaluation of HT1080 tumors in micetreated with placebo, ET-736 and 32.

FIG. 50. Tumor volume evaluation of MDA-MB-231 tumors in mice treatedwith placebo, ET-736 and 32.

FIG. 51. Tumor volume evaluation of H460 tumors in mice treated withplacebo, ET-736 and 32.

FIG. 52. Tumor volume evaluation of H526 tumors in mice treated withplacebo, ET-736 and 32.

FIG. 53. Tumor volume evaluation of H82 tumors in mice treated withplacebo, ET-736 and 32.

FIG. 54. Tumor volume evaluation of A2780 tumors in mice treated withplacebo, ET-736 and 32.

FIG. 55. Tumor volume evaluation of HGC27 tumors in mice treated withplacebo, ET-736 and 32.

FIG. 56. Tumor total diameter evaluation of HT1080 tumors in micetreated with placebo, PM01183 and 35.

FIG. 57. Tumor volume evaluation of MDA-MB-231 tumors in mice treatedwith placebo, PM01183 and 35.

FIG. 58. Tumor volume evaluation of H460 tumors in mice treated withplacebo, PM01183 and 35.

FIG. 59. Tumor volume evaluation of A2780 tumors in mice treated withplacebo, PM01183 and 35.

FIG. 60. Tumor volume evaluation of HGC27 tumors in mice treated withplacebo, PM01183 and 35.

FIG. 61. Tumor volume evaluation of PC-3 tumors in mice treated withplacebo, 12-S and 12-R.

FIG. 62. Tumor volume evaluation of PC-3 tumors in mice treated withplacebo and 4-S.

FIG. 63. Tumor volume evaluation of DU-145 tumors in mice treated withplacebo and 4-S.

FIG. 64. Tumor volume evaluation of 22Rv1 tumors in mice treated withplacebo and 4-S.

FIG. 65. Tumor volume evaluation of PC-3 tumors in mice treated withplacebo and 39-S.

FIG. 66. Tumor volume evaluation of DU-145 tumors in mice treated withplacebo and 39-S.

FIG. 67. Tumor volume evaluation of 22Rv1 tumors in mice treated withplacebo and 39-S.

DETAILED DESCRIPTION OF PREFERRED EMBODIMENTS

The following apply to all aspects of the present invention:

In the compounds of the present invention, the alkyl groups may bebranched or unbranched, and preferably have from 1 to about 12 carbonatoms. One more preferred class of alkyl groups has from 1 to about 6carbon atoms. Even more preferred are alkyl groups having 1, 2, 3 or 4carbon atoms. Methyl, ethyl, n-propyl, isopropyl and butyl, includingn-butyl, isobutyl, sec-butyl and tert-butyl are particularly preferredalkyl groups in the compounds of the present invention.

In the compounds of the present invention, the alkenyl groups may bebranched or unbranched, have one or more double bonds and from 2 toabout 12 carbon atoms. One more preferred class of alkenyl groups hasfrom 2 to about 6 carbon atoms. Even more preferred are alkenyl groupshaving 2, 3 or 4 carbon atoms. Ethenyl, 1-propenyl, 2-propenyl,1-methylethenyl, 1-butenyl, 2-butenyl, and 3-butenyl are particularlypreferred alkenyl groups in the compounds of the present invention.

In the compounds of the present invention, the alkynyl groups may bebranched or unbranched, have one or more triple bonds and from 2 toabout 12 carbon atoms. One more preferred class of alkynyl groups hasfrom 2 to about 6 carbon atoms. Even more preferred are alkynyl groupshaving 2, 3 or 4 carbon atoms.

Suitable aryl groups in the compounds of the present invention includesingle and multiple ring compounds, including multiple ring compoundsthat contain separate and/or fused aryl groups. Typical aryl groupscontain from 1 to 3 separated and/or fused rings and from 6 to about 18carbon ring atoms. Preferably aryl groups contain from 6 to about 10carbon ring atoms. Specially preferred aryl groups included substitutedor unsubstituted phenyl, substituted or unsubstituted naphthyl,substituted or unsubstituted biphenyl, substituted or unsubstitutedphenanthryl and substituted or unsubstituted anthryl.

Suitable heterocyclic groups include heteroaromatic and heteroalicyclicgroups containing from 1 to 3 separated and/or fused rings and from 5 toabout 18 ring atoms. Preferably heteroaromatic and heteroalicyclicgroups contain from 5 to about 10 ring atoms, most preferably 5, 6, or 7ring atoms. Suitable heteroaromatic groups in the compounds of thepresent invention contain one, two or three heteroatoms selected from N,O or S atoms and include, e.g., coumarinyl including 8-coumarinyl,quinolyl including 8-quinolyl, isoquinolyl, pyridyl, pyrazinyl,pyrazolyl, pyrimidinyl, furyl, pyrrolyl, thienyl, thiazolyl,isothiazolyl, triazolyl, tetrazolyl, isoxazolyl, oxazolyl, imidazolyl,indolyl, isoindolyl, indazolyl, indolizinyl, phthalazinyl, pteridyl,purinyl, oxadiazolyl, thiadiazolyl, furazanyl, pyridazinyl, triazinyl,cinnolinyl, benzimidazolyl, benzofuranyl, benzofurazanyl,benzothiophenyl, benzothiazolyl, benzoxazolyl, quinazolinyl,quinoxalinyl, naphthyridinyl and furopyridyl. Suitable heteroalicyclicgroups in the compounds of the present invention contain one, two orthree heteroatoms selected from N, O or S and include, e.g.,pyrrolidinyl, tetrahydrofuranyl, tetrahydrothienyl,tetrahydrothiopyranyl, piperidyl, morpholinyl, thiomorpholinyl,thioxanyl, piperazinyl, azetidinyl, oxetanyl, thietanyl, homopiperidyl,oxepanyl, thiepanyl, oxazepinyl, diazepinyl, thiazepinyl,1,2,3,6-tetrahydropyridyl, 2-pirrolinyl, 3-pyrrolinyl, indolinyl,2H-pyranyl, 4H-pyranyl, dioxanyl, 1,3-dioxolanyl, pyrazolinyl,dithianyl, dithiolanyl, dihydropyranyl, dihydrothienyl, dihydrofuranyl,pyrazolidinyl, imidazolinyl, imidazolidinyl, 3-azabicyclo[3.1.0]hexyl,3-azabicyclo[4.1.0]heptyl, 3H-indolyl, and quinolizinyl.

The groups above mentioned may be substituted at one or more availablepositions by one or more suitable groups such as OR′, ═O, SR′, SOR′,SO₂R′, NO₂, NHR′, NR′R′, ═N—R′, NHCOR′, N(COR′)₂, NHSO₂R′,NR′C(═NR′)NR′R′, CN, halogen, COR′, COOR′, OCOR′, OCONHR′, OCONR′R′,CONHR′, CONR′R′, protected OH, protected amino, protected SH,substituted or unsubstituted C₁-C₁₂ alkyl, substituted or unsubstitutedC₂-C₁₂ alkenyl, substituted or unsubstituted C₂-C₁₂ alkynyl, substitutedor unsubstituted aryl, and substituted or unsubstituted heterocyclicgroup, where each of the R′ groups is independently selected from thegroup consisting of hydrogen, OH, NO₂, NH₂, SH, CN, halogen, COH,COalkyl, CO₂H, substituted or unsubstituted C₁-C₁₂ alkyl, substituted orunsubstituted C₂-C₁₂ alkenyl, substituted or unsubstituted C₂-C₁₂alkynyl, substituted or unsubstituted aryl, and substituted orunsubstituted heterocyclic group. Where such groups are themselvessubstituted, the substituents may be chosen from the foregoing list. Inaddition, where there are more than one R′ groups on a substituent, eachR′ may be the same or different.

In the compounds for the present invention, the halogen substituentsinclude F, Cl, Br, and I.

The terms “pharmaceutically acceptable salt” and “ester” refers to anypharmaceutically acceptable salt or ester which, upon administration tothe patient is capable of providing (directly or indirectly) a compoundas described herein. However, it will be appreciated thatnon-pharmaceutically acceptable salts also fall within the scope of theinvention since those may be useful in the preparation ofpharmaceutically acceptable salts. The preparation of salts can becarried out by methods known in the art.

For instance, pharmaceutically acceptable salts of the compoundsprovided herein are synthesized from the parent compounds, which containa basic or acidic moiety, by conventional chemical methods. Generally,such salts are, for example, prepared by reacting the free acid or baseof these compounds with a stoichiometric amount of the appropriate baseor acid in water or in an organic solvent or in a mixture of both.Generally, nonaqueous media like ether, ethyl acetate, ethanol,2-propanol or acetonitrile are preferred. Examples of the acid additionsalts include mineral acid addition salts such as, for example,hydrochloride, hydrobromide, hydroiodide, sulfate, nitrate, phosphate,and organic acid addition salts such as, for example, acetate,trifluoroacetate, maleate, fumarate, citrate, oxalate, succinate,tartrate, malate, mandelate, methanesulfonate and p-toluenesulfonate.Examples of the alkali addition salts include inorganic salts such as,for example, sodium, potassium, calcium and ammonium salts, and organicalkali salts such as, for example, ethylenediamine, ethanolamine,N,N-dialkylenethanolamine, triethanolamine and basic aminoacids salts.

The compounds of the invention may be in crystalline or amorphous formeither as free compounds or as solvates (e.g. hydrates) and it isintended that all forms are within the scope of the present invention.Methods of solvation are generally known within the art.

Stereoisomerism about the asymmetric carbons with unspecifiedstereochemistry is possible, therefore in such cases the asymmetriccarbons can have (R) or (S) configuration. All diastereomers generatedby a specific configuration of such asymmetric carbons in conjunctionwith the other asymmetric carbons present in the molecule, and mixturesthereof, are considered within the scope of the present invention.Stereoisomerism about the double bond (geometric isomerism) is alsopossible, therefore in some cases the molecule could exist as (E)-isomeror (Z)-isomer. If the molecule contains several double bonds, eachdouble bond will have its own stereoisomerism, that could be the same ordifferent than the stereoisomerism of the other double bonds of themolecule. Furthermore, compounds referred to herein may exist asatropoisomers. The single stereoisomers including diastereoisomers,geometric isomers and atropoisomers of the compounds referred to herein,and mixtures thereof fall within the scope of the present invention.

In addition, compounds referred to herein may exist inisotopically-labelled forms. All pharmaceutically acceptable salts,esters and isotopically labelled forms of the compounds referred toherein, and mixtures thereof, are considered within the scope of thepresent invention.

Protected forms of the compounds disclosed herein are considered withinthe scope of the present invention. Suitable protecting groups are wellknown for the skilled person in the art. A general review of protectinggroups in organic chemistry is provided by Wuts, P G M and Greene T W inProtecting Groups in Organic Synthesis, 4^(th) Ed. Wiley-Interscience,and by Kocienski P J in Protecting Groups, 3^(rd) Ed. Georg ThiemeVerlag. These references provide sections on protecting groups for OH,amino and SH groups. All these references are incorporated by referencein their entirety.

Within the scope of the present invention an OH protecting group isdefined to be the 0-bonded moiety resulting from the protection of theOH through the formation of a suitable protected OH group. Examples ofsuch protected OH groups include ethers, silyl ethers, esters,sulfonates, sulfenates and sulfinates, carbonates, and carbamates. Inthe case of ethers the protecting group for the OH can be selected frommethyl, methoxymethyl, methylthiomethyl,(phenyldimethylsilyl)methoxymethyl, benzyloxymethyl,p-methoxybenzyloxymethyl, [(3,4-dimethoxybenzyl)oxy]methyl,p-nitrobenzyloxymethyl, o-nitrobenzyloxymethyl,[(R)-1-(2-nitrophenyl)ethoxy]methyl, (4-methoxyphenoxy)methyl,guaiacolmethyl, [(p-phenylphenyl)oxy]methyl, t-butoxymethyl,4-pentenyloxymethyl, siloxymethyl, 2-methoxyethoxymethyl,2-cyanoethoxymethyl, bis(2-chloroethoxy)methyl,2,2,2-trichloroethoxymethyl, 2-(trimethylsilyl)ethoxymethyl,menthoxymethyl, O-bis(2-acetoxy-ethoxy)methyl, tetrahydropyranyl,fluorous tetrahydropyranyl, 3-bromotetrahydropyranyl,tetrahydrothiopyranyl, 1-methoxycyclohexyl, 4-methoxytetrahydropyranyl,4-methoxytetrahydrothiopyranyl, 4-methoxytetrahydrothiopyranylS,S-dioxide, 1-[(2-chloro-4-methyl)-phenyl]-4-methoxypiperidin-4-yl,1-(2-fluorophenyl)-4-methoxypiperidin-4-yl,1-(4-chlorophenyl)-4-methoxypiperidin-4-yl, 1,4-dioxan-2-yl,tetrahydropyranyl, tetrahydrothiofuranyl,2,3,3a,4,5,6,7,7a-octahydro-7,8,8-trimethyl-4,7-methanobenzofuran-2-yl,1-ethoxyethyl, 1-(2-chloroethoxy)ethyl, 2-hydroxyethyl, 2-bromoethyl,1-[2-(trimethylsilyl)ethoxy]ethyl, 1-methyl-1-methoxyethyl,1-methyl-1-benzyloxyethyl, 1-methyl-1-benzyloxy-2-fluoroethyl,1-methyl-1-phenoxyethyl, 2,2,2-trichloroethyl,1,1-dianisyl-2,2,2-trichloroethyl,1,1,1,3,3,3-hexafluoro-2-phenylisopropyl, 1-(2-cyanoethoxy)ethyl,2-trimethylsilylethyl, 2-(benzylthio)ethyl, 2-(phenylselenyl)ethyl,t-butyl, cyclohexyl, 1-methyl-1′-cyclopropylmethyl, allyl, prenyl,cinnamyl, 2-phenallyl, propargyl, p-chlorophenyl, p-methoxyphenyl,p-nitrophenyl, 2,4-dinitrophenyl,2,3,5,6-tetrafluoro-4-(trifluoromethyl)phenyl, benzyl, p-methoxybenzyl,3,4-dimethoxybenzyl, 2,6-dimethoxybenzyl, o-nitrobenzyl, p-nitrobenzyl,pentadienylnitrobenzyl, pentadienylnitropiperonyl, halobenzyl,2,6-dichlorobenzyl, 2,4-dichlorobenzyl, 2,6-difluorobenzyl,p-cyanobenzyl, fluorous benzyl, 4-fluorousalkoxybenzyl,trimethylsilylxylyl, p-phenylbenzyl, 2-phenyl-2-propyl,p-acylaminobenzyl, p-azidobenzyl, 4-azido-3-chlorobenzyl,2-trifluoromethylbenzyl, 4-trifluoromethylbenzyl,p-(methylsulfinyl)benzyl, p-siletanylbenzyl, 4-acetoxybenzyl,4-(2-trimethylsilyl)ethoxymethoxybenzyl, 2-naphthylmethyl, 2-picolyl,4-picolyl, 3-methyl-2-picolyl N-oxide, 2-quinolinylmethyl,6-methoxy-2-(4-methylphenyl)-4-quinolinemethyl, 1-pyrenylmethyl,diphenylmethyl, 4-methoxydiphenylmethyl, 4-phenyldiphenylmethyl,p,p′-dinitrobenzhydryl, 5-dibenzosuberyl, triphenylmethyl,tris(4-t-butylphenyl)methyl, α-naphthyldiphenylmethyl,p-methoxyphenyldiphenylmethyl, di(p-methoxyphenyl)phenyl-methyl,tri(p-methoxyphenyl)methyl, 4-(4′-bromophenacyloxy)phenyldiphenylmethyl,4,4′,4″-tris(4,5-dichlorophthalimidophenyl)methyl,4,4′,4″-tris(levulinoyloxyphenyl)methyl,4,4′,4″-tris(benzoyloxyphenyl)methyl,4,4′-dimethoxy-3″-[N-(imidazolylmethyl)]trityl,4,4′-dimethoxy-3″-[N-(imidazolylethyl)carbamoyl]trityl,bis(4-methoxyphenyl)-1′-pyrenylmethyl,4-(17-tetrabenzo[a,c,g,i]fluorenylmethyl)-4,4″-dimethoxytrityl,9-anthryl, 9-(9-phenyl)xanthenyl, 9-phenylthioxanthyl,9-(9-phenyl-10-oxo)anthryl, 1,3-benzodithiolan-2-yl,4,5-bis(ethoxycarbonyl)-[1,3]-dioxolan-2-yl, benzisothiazolylS,S-dioxide. In the case of silyl ethers the protecting group for the OHcan be selected from trimethylsilyl, triethylsilyl, triisopropylsilyl,dimethylisopropylsilyl, diethylisopropylsilyl, dimethylhexylsilyl,2-norbornyldimethylsilyl, t-butyldimethylsilyl, t-butyldiphenylsilyl,tribenzylsilyl, tri-p-xylylsilyl, triphenylsilyl, diphenylmethylsilyl,di-t-butylmethylsilyl, bis(t-butyl)-1-pyrenylmethoxysilyl,tris(trimethylsilyl)silyl, (2-hydroxystyryl)dimethylsilyl,(2-hydroxystyryl)diisopropylsilyl, t-butylmethoxyphenylsilyl,t-butoxydiphenylsilyl, 1,1,3,3-tetraisopropyl-3-[2-(triphenylmethoxy)ethoxy]disiloxane-1-yl, and fluorous silyl. In the case of esters theprotecting group for the OH together with the oxygen atom of theunprotected OH to which it is attached form an ester that can beselected from formate, benzoylformate, acetate, chloroacetate,dichloroacetate, trichloroacetate, trichloroacetamidate,trifluoroacetate, methoxyacetate, triphenylmethoxyacetate,phenoxyacetate, p-chlorophenoxyacetate, phenylacetate, diphenylacetate,3-phenylpropionate, bisfluorous chain type propanoyl, 4-pentenoate,4-oxopentanoate, 4,4-(ethylenedithio)pentanoate,5[3-bis(4-methoxyphenyl)hydro-xymethylphenoxy]levulinate, pivaloate,1-adamantoate, crotonate, 4-methoxycrotonate, benzoate,p-phenylbenzoate, 2,4,6-trimethylbenzoate, 4-bromobenzoate,2,5-difluorobenzoate, p-nitrobenzoate, picolinate, nicotinate,2-(azidomethyl)benzoate, 4-azido-butyrate, (2-azidomethyl)phenylacetate,2-{[(tritylthio)oxy]methyl}benzoate,2-{[(4-methoxytritylthio)oxy]methyl}benzoate,2-{[methyl(tritylthio)amino]methyl}benzoate,2-{[(4-methoxytrityl)thio]methylamino}methyl benzoate,2-(allyloxy)phenylacetate, 2-(prenyloxymethyl)benzoate,6-(levulinyloxymethyl)-3-methoxy-2-nitrobenzoate,6-(levulinyloxymethyl)-3-methoxy-4-nitrobenzoate, 4-benzyloxybutyrate,4-trialkylsilyloxy-butyrate, 4-acetoxy-2,2-dimethylbutyrate,2,2-dimethyl-4-pentenoate, 2-iodobenzoate, 4-nitro-4-methylpentanoate,o-(dibromomethyl)benzoate, 2-formylbenzenesulfonate,4-(methylthio-methoxy)butyrate, 2-(methylthiomethoxymethyl)benzoate,2-(chloroacetoxymethyl)benzoate, 2-[(2-chloroacetoxy)ethyl]benzoate,2-[2-(benzyloxy)ethyl]benzoate,2-[2-(4-methoxybenzyl-oxy)ethyl]benzoate,2,6-dichloro-4-methylphenoxyacetate,2,6-dichloro-4-(1,1,3,3-tetramethylbutyl)phenoxyacetate,2,4-bis(1,1-dimethylpropyl)phenoxyacetate, chlorodiphenyl-acetate,isobutyrate, monosuccinoate, (E)-2-methyl-2-butenoate,o-(methoxycarbonyl)benzoate, α-naphthoate, nitrate, alkylN,N,N′,N′-tetramethylphosphorodiamidate, and 2-chlorobenzoate. In thecase of sulfonates, sulfenates and sulfinates the protecting group forthe OH together with the oxygen atom of the unprotected OH to which itis attached form a sulfonate, sulfenate or sulfinates that can beselected from sulfate, allylsulfonate, methanesulfonate,benzylsulfonate, tosylate, 2[(4-nitrophenyl)ethyl]sulfonate,2-trifluoromethylbenzenesulfonate, 4-monomethoxytritylsulfenate, alkyl2,4-dinitrophenylsulfenate,2,2,5,5-tetramethylpyrrolidin-3-one-1-sulfinate, anddimethylphosphinothioyl. In the case of carbonates the protecting groupfor the OH together with the oxygen atom of the unprotected OH to whichit is attached form a carbonate that can be selected from methylcarbonate, methoxymethyl carbonate, 9-fluorenylmethyl carbonate, ethylcarbonate, bromoethyl carbonate, 2-(methylthiomethoxy)ethyl carbonate,2,2,2-trichloroethyl carbonate, 1,1-dimethyl-2,2,2-trichloroethylcarbonate, 2-(trimethylsilyl)ethyl carbonate,2-[dimethyl(2-naphthylmethyl)silyl]ethyl carbonate,2-(phenylsulfonyl)ethyl carbonate, 2-(triphenylphosphonio)ethylcarbonate, cis-[4-[[(methoxytrityl)sulfenyl]oxy]tetrahydrofuran-3-yl]oxycarbonate, isobutyl carbonate, t-butyl carbonate, vinyl carbonate, allylcarbonate, cinnamyl carbonate, propargyl carbonate, p-chlorophenylcarbonate, p-nitrophenyl carbonate, 4-ethoxy-1-naphthyl carbonate,6-bromo-7-hydroxycoumarin-4-ylmethyl carbonate, benzyl carbonate,o-nitrobenzyl carbonate, p-nitrobenzyl carbonate, p-methoxybenzylcarbonate, 3,4-dimethoxybenzyl carbonate, anthraquinon-2-ylmethylcarbonate, 2-dansylethyl carbonate, 2-(4-nitrophenyl)ethyl carbonate,2-(2,4-dinitrophenyl)ethyl carbonate, 2-(2-nitrophenyl)propyl carbonate,2-(3,4-methylenedioxy-6-nitrophenyl)propyl carbonate,2-cyano-1-phenylethyl carbonate, 2-(2-pyridyl)amino-1-phenylethylcarbonate, 2-[N-methyl-N-(2-pyridyl)]amino-1-phenylethyl carbonate,phenacyl carbonate, 3′,5′-dimethoxybenzoin carbonate, methyldithiocarbonate, and S-benzyl thiocarbonate. And in the case ofcarbamates the protecting group for OH together with the oxygen atom ofthe unprotected OH to which it is attached forms a carbamate that can beselected from dimethyl thiocarbamate, N-phenyl carbamate, andN-methyl-N-(o-nitrophenyl) carbamate.

Within the scope of the present invention an amino protecting group isdefined to be the N-bonded moiety resulting from the protection of theamino group through the formation of a suitable protected amino group.Examples of protected amino groups include carbamates, ureas, amides,heterocyclic systems, N-alkyl amines, N-alkenyl amines, N-alkynylamines, N-aryl amines, imines, enamines, N-metal derivatives, N—Nderivatives, N—P derivatives, N—Si derivatives, and N—S derivatives. Inthe case of carbamates the protecting group for the amino group togetherwith the amino group to which it is attached form a carbamate that canbe selected from methyl carbamate, ethyl carbamate, 9-fluorenylmethylcarbamate, 2,6-di-t-butyl-9-fluorenylmethyl carbamate,2,7-bis(trimethylsilyl)fluorenylmethyl carbamate,9-(2-sulfo)fluorenylmethyl carbamate, 9-(2,7-dibromo)fluorenylmethylcarbamate, 17-tetrabenzo[a,c,g,i]fluorenylmethyl carbamate,2-chloro-3-indenylmethyl carbamate, benz[f]inden-3-ylmethyl carbamate,1,1-dioxobenzo[b]-thiophene-2-ylmethyl carbamate,2-methylsulfonyl-3-phenyl-1-prop-2-enyl carbamate,2,7-di-t-butyl-[9,(10,10-dioxo-10,10,10,10-tetrahydrothioxanthyl)]methylcarbamate, 2,2,2-trichloroethyl carbamate, 2-trimethylsilylethylcarbamate, (2-phenyl-2-trimethylsilyl)ethyl carbamate, 2-phenylethylcarbamate, 2-chloroethyl carbamate, 1,1-dimethyl-2-haloethyl carbamate,1,1-dimethyl-2,2-dibromoethyl carbamate,1,1-dimethyl-2,2,2-trichloroethyl carbamate, 2-(2′-pyridyl)ethylcarbamate, 2-(4′-pyridyl)ethyl carbamate, 2,2-bis(4′-nitrophenyl)ethylcarbamate, 2[(2-nitrophenyl)dithio]-1-phenylethyl carbamate,2-(N,N-dicyclohexylcarboxamido)ethyl carbamate, t-butyl carbamate,fluorous BOC carbamate, 1-adamantyl carbamate, 2-adamantyl carbamate,1-(1-adamantyl)-1-methylethyl carbamate, 1-methyl-1-(4-byphenylyl)ethylcarbamate, 1-(3,5-di-t-butylphenyl)-1-methylethyl carbamate,triisopropylsilyloxy carbamate, vinyl carbamate, allyl carbamate, prenylcarbamate, 1-isopropylallyl carbamate, cinnamyl carbamate,4-nitrocinnamyl carbamate, 3-(3′-pyridyl)prop-2-enyl carbamate,hexadienyl carbamate, propargyl carbamate, 1,4-but-2-ynyl biscarbamate,8-quinolyl carbamate, N-hydroxypiperidinyl carbamate, alkyldithiocarbamate, benzyl carbamate, 3,5-di-t-butylbenzyl carbamate,p-methoxybenzyl carbamate, p-nitrobenzyl carbamate, p-bromobenzylcarbamate, p-chlorobenzyl carbamate, 2,4-dichlorobenzyl carbamate,4-methylsulfinylbenzyl carbamate, 4-trifluoromethylbenzyl carbamate,fluorous benzyl carbamate, 2-naphthylmethyl carbamate, 9-anthrylmethylcarbamate, diphenylmethyl carbamate, 4-phenylacetoxybenzyl carbamate,4-azidobenzyl carbamate, 4-azido-methoxybenzyl carbamate,m-chloro-p-acyloxybenzyl carbamate, p-(dihydroxyboryl)-benzyl carbamate,5-benzisoxazolylmethyl carbamate, 2-(trifluoromethyl)-6-chromonylmethylcarbamate, 2-methylthioethyl carbamate, 2-methylsulfonylethyl carbamate,2-(p-toluenesulfonyl)ethyl carbamate, 2-(4-nitrophenylsulfonyl)ethylcarbamate, 2-(2,4-dinitrophenylsulfonyl)ethyl carbamate,2-(4-trifluoromethylphenylsulfonyl)ethyl carbamate,[2-(1,3-dithianyl)]methyl carbamate, 2-phosphonioethyl carbamate,2-[phenyl(methyl)sulfonio]ethyl carbamate,1-methyl-1-(triphenylphosphonio)ethyl carbamate,1,1-dimethyl-2-cyanoethyl carbamate, 2-dansylethyl carbamate,2-(4-nitrophenyl)ethyl carbamate, 4-methylthiophenyl carbamate,2,4-dimethylthiophenyl carbamate, m-nitrophenyl carbamate,3,5-dimethoxybenzyl carbamate, 1-methyl-1-(3,5-dimethoxyphenyl)ethylcarbamate, α-methylnitropiperonyl carbamate, o-nitrobenzyl carbamate,3,4-dimethoxy-6-nitrobenzyl carbamate, phenyl(o-nitrophenyl)methylcarbamate, 2-nitrophenylethyl carbamate, 6-nitroveratryl carbamate,4-methoxyphenacyl carbamate, 3′,5′-dimethoxybenzoin carbamate,9-xanthenylmethyl carbamate, N-methyl-N-(o-nitrophenyl) carbamate,t-amyl carbamate, 1-methylcyclobutyl carbamate, 1-methylcyclohexylcarbamate, 1-methyl-1-cyclopropylmethyl carbamate, cyclobutyl carbamate,cyclopentyl carbamate, cyclohexyl carbamate, isobutyl carbamate,isobornyl carbamate, cyclopropylmethyl carbamate, p-decyloxybenzylcarbamate, diisopropylmethyl carbamate, 2,2-dimethoxy-carbonylvinylcarbamate, o-(N,N-dimethylcarboxamido)benzyl carbamate,1,1-dimethyl-3-(N,N-dimethyl-carboxamido)propyl carbamate, butynylcarbamate, 1,1-dimethylpropynyl carbamate, 2-iodoethyl carbamate,1-methyl-1-(4′-pyridyl)ethyl carbamate,1-methyl-1-(p-phenylazophenyl)ethyl carbamate,p-(p′-methoxyphenylazo)benzyl carbamate, p-(phenylazo)benzyl carbamate,2,4,6-trimethylbenzyl carbamate, isonicotinyl carbamate,4-(trimethyl-ammonium)benzyl carbamate, p-cyanobenzyl carbamate,di(2-pyridyl)methyl carbamate, 2-furanylmethyl carbamate, phenylcarbamate, 2,4,6-tri-t-butylphenyl carbamate, 1-methyl-1-phenylethylcarbamate, and S-benzyl thiocarbamate. In the case of ureas theprotecting groups for the amino group can be selected fromphenothiazinyl-(10)-carbonyl, N′-p-toluenesulfonylaminocarbonyl,N′-phenylaminothiocarbonyl, 4-hydroxyphenylaminocarbonyl,3-hydroxytryptaminocarbonyl, and N′-phenylaminothiocarbonyl. In the caseof amides the protecting group for the amino together with the aminogroup to which it is attached form an amide that can be selected fromformamide, acetamide, chloroacetamide, trichloroacetamide,trifluoroacetamide, phenylacetamide, 3-phenylpropanamide,pent-4-enamide, picolinamide, 3-pyridylcarboxamide,N-benzoylphenylalanyl amide, benzamide, p-phenylbenzamide,o-nitrophenylacetamide, 2,2-dimethyl-2-(o-nitrophenyl)acetamide,o-nitrophenoxyacetamide, 3-(o-nitrophenyl)propanamide,2-methyl-2-(o-nitrophenoxy)propanamide, 3-methyl-3-nitrobutanamide,o-nitrocinnamide, o-nitrobenzamide,3-(4-t-butyl-2,6-dinitrophenyl)-2,2-dimethylpropanamide,o-(benzoyloxyme-thyl)benzamide, 2-(acetoxymethyl)benzamide,2-[(t-butyldiphenylsiloxy)methyl]benzamide,3-(3′,6′-dioxo-2′,4′,5′-trimethylcyclohexa-1′,4′-diene)-3,3-dimethylpropionamide,o-hydroxy-trans-cinnamide, 2-methyl-2-(o-phenylazophenoxy)propanamide,4-chlorobutanamide, aceto-acetamide, 3-(p-hydroxyphenyl)propanamide,(N-dithiobenzyloxycarbonylamino)acetamide, and N-acetylmethionine amide.In the case of heterocyclic systems the protecting group for the aminogroup together with the amino group to which it is attached form aheterocyclic system that can be selected from4,5-diphenyl-3-oxazolin-2-one, N-phthalimide, N-dichlorophthalimide,N-tetrachlorophthalimide, N-4-nitrophthalimide, N-thiodiglycoloyl,N-dithiasuccinimide, N-2,3-diphenylmaleimide, N-2,3-dimethylmaleimide,N-2,5-dimethylpyrrole, N-2,5-bis(triisopropylsiloxy)pyrrole,N-1,1,4,4-tetramethyldisilylazacyclopentane adduct,N-1,1,3,3-tetramethyl-1,3-disilaisoindoline, N-diphenylsilyldiethylene,N-5-substituted-1,3-dimethyl-1,3,5-triazacyclohexan-2-one,N-5-substituted-1,3-benzyl-1,3,5-triazacyclohexan-2-one, 1-substituted3,5-dinitro-4-pyridone, and 1,3,5-dioxazine. In the case of N-alkyl,N-alkenyl, N-alkynyl or N-aryl amines the protecting group for the aminogroup can be selected from N-methyl, N-t-butyl, N-allyl, N-prenyl,N-cinnamyl, N-phenylallyl, N-propargyl, N-methoxymethyl,N-[2-(trimethylsilyl)ethoxy]methyl, N-3-acetoxypropyl, N-cyanomethyl,N-2-azanorbornenes, N-benzyl, N-4-methoxybenzyl, N-2,4-dimethoxybenzyl,N-2-hydroxybenzyl, N-ferrocenylmethyl, N-2,4-dinitrophenyl,o-methoxyphenyl, p-methoxyphenyl, N-9-phenylfluorenyl, N-fluorenyl,N-2-picolylamine N′-oxide, N-7-methoxycoumar-4-ylmethyl,N-diphenylmethyl, N-bis(4-methoxyphenyl)methyl, N-5-dibenzosuberyl,N-triphenylmethyl, N-(4-methylphenyl)diphenylmethyl, andN-(4-methoxyphenyl)diphenylmethyl. In the case of imines the protectinggroup for the amino group can be selected fromN-1,1-dimethylthiomethylene, N-benzylidene, N-p-methoxybenzylidene,N-diphenylmethylene, N-[2-pyridyl)mesityl]methylene,N—(N,N-dimethylaminomethylene), N—(N,N-dibenzylaminomethylene),N—(N-t-butylaminomethylene), N,N-isopropylidene, N-p-nitrobenzylidene,N-salicylidene, N-5-chlorosalicylidene,N-(5-chloro-2-hydroxyphenyl)phenylmethylene, N-cyclohexylidene, andN-t-butylidene. In the case of enamines the protecting group for theamino group can be selected from N-(5,5-dimethyl-3-oxo-1-cyclohexenyl),N-2,7-dichloro-9-fluorenylmethylene,N-1-(4,4-dimethyl-2,6-dioxocyclohexylidene)ethyl,N-(1,3-dimethyl-2,4,6-(1H,3H,5H)-trioxopyrimidine-5-ylidene)-methyl,N-4,4,4-trifluoro-3-oxo-1-butenyl, andN-(1-isopropyl-4-nitro-2-oxo-3-pyrrolin-3-yl). In the case of N-metalderivatives the protecting group for the amino group can be selectedfrom N-borane, N-diphenylborinic ester, N-diethylborinic ester,N-9-borabicyclononane, N-difluoroborinic ester, and3,5-bis(trifluoromethyl)phenylboronic acid; and also includingN-phenyl(pentacarbonylchromium)carbenyl,N-phenyl(pentacarbonyl-tungsten)carbenyl,N-methyl(pentacarbonylchromium)carbenyl,N-methyl(pentacarbonyltungsten)carbenyl, N-copper chelate, N-zincchelate, and a 18-crown-6-derivative. In the case of N—N derivatives theprotecting group for the amino group together with the amino group towhich it is attached form a N—N derivative that can be selected fromN-nitroamino, N-nitrosoamino, amine N-oxide, azide, triazene derivative,and N-trimethylsilylmethyl-N-benzylhydrazine. In the case of N—Pderivatives the protected group for the amino group together with theamino group to which it is attached form a N—P derivative that can beselected from diphenylphosphinamide, dimethylthiophosphinamide,diphenylthiophosphinamide, dialkyl phosphoramidate, dibenzylphosphoramidate, diphenyl phosphoramidate, andiminotriphenylphosphorane. In the case of N—Si derivatives theprotecting group for the NH₂ can be selected from t-butyldiphenylsilyland triphenylsilyl. In the case of N—S derivatives the protected aminogroup can be selected from N-sulfenyl or N-sulfonyl derivatives. TheN-sulfenyl derivatives can be selected from benzenesulfenamide,2-nitrobenzenesulfenamide, 2,4-dinitrobenzenesulfenamide,pentachlorobenzenesulfenamide, 2-nitro-4-methoxybenzenesulfenamide,triphenylmethylsulfe-namide,1-(2,2,2-trifluoro-1,1-diphenyl)ethylsulfenamide, andN-3-nitro-2-pyridinesulfenamide. The N-sulfonyl derivatives can beselected from methanesulfonamide, trifluoromethanesulfonamide,t-butylsulfonamide, benzylsulfonamide, 2-(trimethylsilyl)ethanesulfonamide, p-toluenesulfonamide, benzenesulfonamide,o-anisylsulfonamide, 2-nitrobenzenesulfonamide,4-nitrobenzenesulfonamide, 2,4-dinitrobenzenesulfonamide,2-naphthalenesulfonamide,4-(4′,8′-dimethoxynaphthylmethyl)benzenesulfonamide,2-(4-methylphenyl)-6-methoxy-4-methylsulfonamide,9-anthracenesulfonamide, pyridine-2-sulfonamide,benzothiazole-2-sulfonamide, phenacylsulfonamide,2,3,6-trimethyl-4-methoxybenzenesulfonamide,2,4,6-trimethoxybenzenesulfonamide,2,6-dimethyl-4-methoxybenzenesulfonamide, pentamethylbenzenesulfonamide,2,3,5,6-tetramethyl-4-methoxybenzenesulfonamide,4-methoxybenzenesulfonamide, 2,4,6-trimethylbenzenesulfonamide,2,6-dimethoxy-4-methylbenzenesulfonamide,3-methoxy-4-t-butylbenzenesulfonamide, and2,2,5,7,8-pentamethylchroman-6-sulfonamide.

Within the scope of the present invention a protecting group for SH isdefined to be the S-bonded moiety resulting from the protection of theSH group through the formation of a suitable a protected SH group.Examples of such protected SH groups include thioethers, disulfides,silyl thioethers, thioesters, thiocarbonates, and thiocarbamates. In thecase of thioethers the protecting group for the SH can be selected fromS-alkyl, S-benzyl, S-p-methoxybenzyl, S-o-hydroxybenzyl,S-p-hydroxybenzyl, S-o-acetoxybenzyl, S-p-acetoxybenzyl,S-p-nitrobenzyl, S-o-nitrobenzyl, S-2,4,6-trimethylbenzyl,S-2,4,6,-trimethoxybenzyl, S-4-picolyl, S-2-picolyl-N-oxide,S-2-quinolinylmethyl, S-9-anthrylmethyl, S-9-fluorenylmethyl,S-xanthenyl, S-ferrocenylmethyl, S-diphenylmethyl,S-bis(4-methoxyphenyl)methyl, S-5-dibenzosuberyl, S-triphenylmethyl,4-methoxytrityl, S-diphenyl-4-pyridylmethyl, S-phenyl,S-2,4-dinitrophenyl, S-2-quinolyl, S-t-butyl, S-1-adamantyl,S-methoxymethyl, S-isobutoxymethyl, S-benzyloxymethyl, S-1-ethoxyethyl,S-2-tetrahydropyranyl, S-benzylthiomethyl, S-phenylthiomethyl,S-acetamidomethyl (Acm), 5-trimethylacetamidomethyl, S-benzamidomethyl,S-allyloxycarbonylaminomethyl,tetrafluoro-4-(N-piperidino)-phenyl-N-allyloxycarbonylaminomethyl,S-phthalimidomethyl, S-phenylacetamidomethyl, S-acetylmethyl,S-carboxymethyl, 5-cyanomethyl, S-(2-nitro-1-phenyl)ethyl,S-2-(2,4-dinitrophenyl)ethyl, S-2-(4′-pyridyl)ethyl, S-2-cyanoethyl,S-2-(trimethylsilyl)ethyl, S-2,2-bis(carboethoxy)ethyl,S-(1-m-nitrophenyl-2-benzoyl)ethyl, S-2-phenylsulfonylethyl,S-1-(4-methylphenylsulfonyl)-2-methylprop-2-yl, and S-p-hydroxyphenacyl.In the case of disulfides the protected SH group can be selected fromS-ethyl disulfide, S-t-butyl disulfide, S-2-nitrophenyl disulfide,S-2,4-dinitrophenyl disulfide, S-2-phenylazophenyl disulfide,S-2-carboxyphenyl disulfide, and S-3-nitro-2-pyridyl disulfide. In thecase of silyl thioethers the protecting group for the SH can be selectedfrom the list of groups that was listed above for the protection of OHwith silyl ethers. In the case of thioesters the protecting group forthe SH can be selected from S-acetyl, S-benzoyl, S-2-methoxyisobutyryl,S-trifluoroacetyl,S—N-[[p-biphenylyl)-isopropyloxy]carbonyl]-N-methyl-γ-aminothiobutyrate,and S—N-(t-butoxycarbonyl)-N-methyl-γ-aminothiobutyrate. In the case ofthiocarbonate protecting group for the SH can be selected fromS-2,2,2-trichloroethoxycarbonyl, S-t-butoxycarbonyl,S-benzyloxycarbonyl, S-p-methoxybenzyloxycarbonyl, andS-fluorenylmethylcarbonyl. In the case of thiocarbamate the protected SHgroup can be selected from S—(N-ethylcarbamate) andS—(N-methoxymethylcarbamate).

The mention of these groups should not be interpreted as a limitation ofthe scope of the invention, since they have been mentioned as a mereillustration of protecting groups for OH, amino and SH groups, butfurther groups having said function may be known by the skilled personin the art, and they are to be understood to be also encompassed by thepresent invention.

To provide a more concise description, some of the quantitativeexpressions given herein are not qualified with the term “about”. It isunderstood that, whether the term “about” is used explicitly or not,every quantity given herein is meant to refer to the actual given value,and it is also meant to refer to the approximation to such given valuethat would reasonably be inferred based on the ordinary skill in theart, including equivalents and approximations due to the experimentaland/or measurement conditions for such given value.

In an embodiment, the compound may be a compound of formula I or apharmaceutically acceptable salt or ester thereof:

wherein:

X is —NH— or —O—; R₁ is —OH or —CN;

R₂ is a —C(═O)R^(a) group;R₃ is hydrogen or a —OR^(b) group;R₄ is selected from hydrogen, —CH₂OH, —CH₂OC(═O)R^(c), —CH₂NH₂, and—CH₂NHProt^(NH);R^(a) is selected from hydrogen, substituted or unsubstituted C₁-C₁₂alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl, and substituted orunsubstituted C₂-C₁₂ alkynyl;R^(b) is selected from substituted or unsubstituted C₁-C₁₂ alkyl,substituted or unsubstituted C₂-C₁₂ alkenyl, and substituted orunsubstituted C₂-C₁₂ alkynyl;R^(c) is selected from substituted or unsubstituted C₁-C₁₂ alkyl,substituted or unsubstituted C₂-C₁₂ alkenyl, and substituted orunsubstituted C₂-C₁₂ alkynyl; andProt^(NH) is a protecting group for amino, with the proviso that when R₄is hydrogen then X is —O—.

In a further embodiment, the compound of formula I may be a compound offormula IC, or a pharmaceutically acceptable salt or ester thereof:

wherein:

-   -   X is —NH—;    -   R₁ is —OH or —CN;    -   R₂ is a —C(═O)R^(a) group;    -   R₃ is hydrogen or a —OR^(b) group;    -   R₄ is selected from —CH₂OH, —CH₂O—(C═O)R^(c), —CH₂NH₂ and        —CH₂NHProt^(NH);    -   R^(a) is selected from hydrogen, substituted or unsubstituted        C₁-C₁₂ alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl, and        substituted or unsubstituted C₂-C₁₂ alkynyl;    -   R^(b) is selected from substituted or unsubstituted C₁-C₁₂        alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl and        substituted or unsubstituted C₂-C₁₂ alkynyl;    -   R^(c) is selected from substituted or unsubstituted C₁-C₁₂        alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl, and        substituted or unsubstituted C₂-C₁₂ alkynyl; and    -   Prot^(NH) is a protecting group for amino.

In a yet further embodiment, the compound of formula I may be a compoundof formula ID, or a pharmaceutically acceptable salt or ester thereof:

wherein:

-   -   X is —O—;    -   R₁ is —OH or —CN;    -   R₂ is a —C(═O)R^(a) group;    -   R₃ is hydrogen or a —OR^(b) group;    -   R₄ is selected from hydrogen, —CH₂OH, —CH₂O—(C═O)R^(c), —CH₂NH₂        and —CH₂NHProt^(NH);    -   R^(a) is selected from hydrogen, substituted or unsubstituted        C₁-C₁₂ alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl, and        substituted or unsubstituted C₂-C₁₂ alkynyl;    -   R^(b) is selected from substituted or unsubstituted C₁-C₁₂        alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl and        substituted or unsubstituted C₂-C₁₂ alkynyl;    -   R^(c) is selected from substituted or unsubstituted C₁-C₁₂        alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl, and        substituted or unsubstituted C₂-C₁₂ alkynyl; and    -   Prot^(NH) is a protecting group for amino.

In a yet further embodiment, the compound of formula I may be a compoundof formula IE, or a pharmaceutically acceptable salt or ester thereof:

wherein:

-   -   X is —NH— or —O—;    -   R₁ is —OH or —CN;    -   R₂ is a —C(═O)R^(a) group;    -   R₃ is hydrogen or a —OR^(b) group;    -   R₄ is selected from —CH₂NH₂ and —CH₂NHProt^(NH);    -   R^(a) is selected from hydrogen, substituted or unsubstituted        C₁-C₁₂ alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl, and        substituted or unsubstituted C₂-C₁₂ alkynyl;    -   R^(b) is selected from substituted or unsubstituted C₁-C₁₂        alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl and        substituted or unsubstituted C₂-C₁₂ alkynyl; and    -   Prot^(NH) is a protecting group for amino.

In a yet further embodiment, the compound of formula I may be a compoundof formula IA or a pharmaceutically acceptable salt or ester thereof:

wherein:

-   -   X is —NH— or —O—;    -   R₁ is —OH or —CN;    -   R₂ is a —C(═O)R^(a) group;    -   R₃ is hydrogen;    -   R₄ is selected from hydrogen, —CH₂OH, —CH₂O—(C═O)R^(c), —CH₂NH₂        and —CH₂NHProt^(NH);    -   R^(a) is selected from hydrogen, substituted or unsubstituted        C₁-C₁₂ alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl, and        substituted or unsubstituted C₂-C₁₂ alkynyl;    -   R^(c) is selected from substituted or unsubstituted C₁-C₁₂        alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl, and        substituted or unsubstituted C₂-C₁₂ alkynyl; and    -   Prot^(NH) is a protecting group for amino;    -   with the proviso that when R₄ is hydrogen then X is —O—.

In a yet further embodiment, the compound of formula I may be a compoundof formula IB or a pharmaceutically acceptable salt or ester thereof:

wherein:

-   -   X is —NH— or —O—;    -   R₁ is —OH or —CN;    -   R₂ is a —C(═O)R^(a) group;    -   R₃ is a —OR^(b) group;    -   R₄ is selected from hydrogen, —CH₂OH, —CH₂O—(C═O)R^(c), —CH₂NH₂        and —CH₂NHProt^(NH);    -   R^(a) is selected from hydrogen, substituted or unsubstituted        C₁-C₁₂ alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl, and        substituted or unsubstituted C₂-C₁₂ alkynyl;    -   R^(b) is selected from substituted or unsubstituted C₁-C₁₂        alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl and        substituted or unsubstituted C₂-C₁₂ alkynyl;    -   R^(c) is selected from substituted or unsubstituted C₁-C₁₂        alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl, and        substituted or unsubstituted C₂-C₁₂ alkynyl; and    -   Prot^(NH) is a protecting group for amino;    -   with the proviso that when R₄ is hydrogen then X is —O—.

In a yet further embodiment, the compound of formula I may be a compoundof formula IF or a pharmaceutically acceptable salt or ester thereof:

wherein:

X is —NH— or —O—; R₁ is —OH;

R₂ is a —C(═O)R^(a) group;R₃ is hydrogen or a —OR^(b) group;R₄ is selected from hydrogen, —CH₂OH, —CH₂OC(═O)R^(c), —CH₂NH₂, and—CH₂NHProt^(NH);R^(a) is selected from hydrogen, substituted or unsubstituted C₁-C₁₂alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl, and substituted orunsubstituted C₂-C₁₂ alkynyl;R^(b) is selected from substituted or unsubstituted C₁-C₁₂ alkyl,substituted or unsubstituted C₂-C₁₂ alkenyl, and substituted orunsubstituted C₂-C₁₂ alkynyl;R^(c) is selected from substituted or unsubstituted C₁-C₁₂ alkyl,substituted or unsubstituted C₂-C₁₂ alkenyl, and substituted orunsubstituted C₂-C₁₂ alkynyl; andProt^(NH) is a protecting group for amino,with the proviso that when R₄ is hydrogen then X is —O—.

In a yet further embodiment, the compound of formula I may be a compoundof formula IG or a pharmaceutically acceptable salt or ester thereof:

wherein:

X is —NH— or —O—; R₁ is —OH or —CN;

R₂ is acetyl;R₃ is hydrogen or a —OR^(b) group;R₄ is selected from hydrogen, —CH₂OH, —CH₂OC(═O)R^(c), —CH₂NH₂, and—CH₂NHProt^(NH);R^(b) is selected from substituted or unsubstituted C₁-C₁₂ alkyl,substituted or unsubstituted C₂-C₁₂ alkenyl, and substituted orunsubstituted C₂-C₁₂ alkynyl;R^(c) is selected from substituted or unsubstituted C₁-C₁₂ alkyl,substituted or unsubstituted C₂-C₁₂ alkenyl, and substituted orunsubstituted C₂-C₁₂ alkynyl; andProt^(NH) is a protecting group for amino,with the proviso that when R₄ is hydrogen then X is —O—.

Preferred compounds of the compounds of formula I, IA, IB, IC, ID, IE,IF, or IG, are those having general formula a or b, or apharmaceutically acceptable salt or ester thereof:

Note where the compounds have general formula a or b, R₄ may not behydrogen.

Preferred compounds of the compounds of formula I, IA, IB, ID, IF, or IGmay be those having formula c or a pharmaceutically acceptable salt orester thereof:

wherein:

R₁ is —OH or —CN;

R₂ is a —C(═O)R^(a) group;R₃ is hydrogen or a —OR^(b) group;R^(a) is selected from hydrogen, substituted or unsubstituted C₁-C₁₂alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl, and substituted orunsubstituted C₂-C₁₂ alkynyl; andR^(b) is selected from substituted or unsubstituted C₁-C₁₂ alkyl,substituted or unsubstituted C₂-C₁₂ alkenyl, and substituted orunsubstituted C₂-C₁₂ alkynyl.

Preferred compounds include compounds of general formula I, IA, IB, IE,IF, IG, Ia, IAa, IBa, IEa, IFa, IGa, Ib, IAb, IBb, IEb, IFb, and IGb,wherein:

X is —NH—;

and R₁; R₂; R₃; R₄; R^(a); R^(b); R^(c); and Prot^(NH) are as defined asabove.

Preferred compounds include compounds of general formula I, IA, IB, IE,IF, IG, Ia, IAa, IBa, IEa, IFa, IGa, Ib, IAb, IBb, IEb, IFb, and IGb,wherein:

X is —O—;

and R₁; R₂; R₃; R₄; R^(a); R^(b); R^(c); and Prot^(NH) are as defined asabove.

Further preferred compounds include compounds of general formula I, IA,IB, IC, ID, IE, IG, Ia, IAa, IBa, ICa, IDa, IEa, IGa, Ib, IAb, IBb, ICb,IDb, IEb, and IGb, wherein:

R₁ is —OH;

and X; R₂; R₃; R₄; R^(a); R^(b); R^(c); and Prot^(NH) are as defined asabove.

Further preferred compounds include compounds of general formula I, IA,IB, IC, ID, IE, IF, Ia, IAa, IBa, ICa, IDa, IEa, IFa, Ib, IAb, IBb, ICb,IDb, IEb, and IFb, wherein:

R₂ is a —C(═O)R^(a) group where R^(a) is a substituted or unsubstitutedC₁-C₆ alkyl. Particularly preferred R^(a) is selected from substitutedor unsubstituted methyl, substituted or unsubstituted ethyl, substitutedor unsubstituted n-propyl, substituted or unsubstituted isopropyl,substituted or unsubstituted n-butyl, substituted or unsubstitutedisobutyl, substituted or unsubstituted sec-butyl and substituted orunsubstituted tert-butyl. Most preferred R₂ is acetyl;and X; R₁; R₃; R₄; R^(b); R^(c); and Prot^(NH) are as defined as above.

Further preferred compounds include compounds of general formula I, IB,IC, ID, IE, IF, IG, Ia, IBa, ICa, IDa, IEa, IFa, IGa, Ib, IBb, ICb, IDb,IEb, IFb, and IGb, wherein:

R₃ is hydrogen or a —OR^(b) group for compounds of formula I, IC, ID,IE, IF, IG, Ia, ICa, IDa, IEa, IFa, IGa, Ib, ICb, IDb, IEb, IFb, or IGb;and R₃ is a —OR^(b) group for compounds of formula IB, IBa or IBb; whereR^(b) is a substituted or unsubstituted C₁-C₆ alkyl. Particularlypreferred R^(b) is selected from substituted or unsubstituted methyl,substituted or unsubstituted ethyl, substituted or unsubstitutedn-propyl, substituted or unsubstituted isopropyl, substituted orunsubstituted n-butyl, substituted or unsubstituted isobutyl,substituted or unsubstituted sec-butyl and substituted or unsubstitutedtert-butyl. More preferred R₃ are hydrogen and methoxy, being hydrogenthe most preferred R₃ group;and X; R₁; R₂; R₄; R^(a); R^(c); and Prot^(NH) are as defined as above.

Further preferred compounds include compounds of general formula I, IA,IB, IC, ID, IE, IF, IG, Ia, IAa, IBa, ICa, IDa, IEa, IFa, IGa, Ib, IAb,IBb, ICb, IDb, IEb, IFb, and IGb, wherein:

R₄ is selected from —CH₂OH, —CH₂OC(═O)R^(c), —CH₂NH₂, and—CH₂NHProt^(NH) for compounds of formula I, IA, IB, IC, ID, IF, IG, Ia,IAa, IBa, ICa, IDa, IFa, IGa, Ib, IAb, IBb, ICb, IDb, IFb, or IGb; andR₄ is selected from —CH₂NH₂, and —CH₂NHProt^(NH) for compounds offormula IE, IEa or IEb; where R^(c) is a substituted or unsubstitutedC₁-C₆ alkyl. Particularly preferred R^(c) is selected from substitutedor unsubstituted methyl, substituted or unsubstituted ethyl, substitutedor unsubstituted n-propyl, substituted or unsubstituted isopropyl,substituted or unsubstituted n-butyl, substituted or unsubstitutedisobutyl, substituted or unsubstituted sec-butyl, and substituted orunsubstituted tert-butyl. Most preferred R^(c) is methyl. More preferredR₄ is selected from —CH₂OH and —CH₂NH₂. More preferably, R₄ may be—CH₂NH₂. Most preferred R₄ is —CH₂OH;and X; R₁; R₂; R₃; R^(a); and R^(b) are as defined as above.

Further preferred compounds include compounds of general formula I, IA,IB, IC, IE, IF, IG, Ia, IAa, IBa, ICa, IEa, IFa, IGa, Ib, IAb, IBb, ICb,IEb, IFb, and IGb, wherein:

X is —NH—; R₁ is —OH;

and R₂; R₃; R₄; R^(a); R^(b); R^(c); and Prot^(NH) are as defined asabove.

Further preferred compounds include compounds of general formula I, IA,IB, IC, IE, IF, IG, Ia, IAa, IBa, ICa, IEa, IFa, IGa, Ib, IAb, IBb, ICb,IEb, IFb, and IGb, wherein:

X is —NH—;

R₂ is a —C(═O)R^(a) for compounds of formula I, IA, IB, IC, IE, IF, Ia,IAa, IBa, ICa, IEa, IFa, Ib, IAb, IBb, ICb, IEb, or IFb; and R₂ isacetyl for compounds of formula IG, IGa or IGb; where R^(a) is asubstituted or unsubstituted C₁-C₆ alkyl. Particularly preferred R^(a)is selected from substituted or unsubstituted methyl, substituted orunsubstituted ethyl, substituted or unsubstituted n-propyl, substitutedor unsubstituted isopropyl, substituted or unsubstituted n-butyl,substituted or unsubstituted isobutyl, substituted or unsubstitutedsec-butyl and substituted or unsubstituted tert-butyl. Most preferred R₂is acetyl;and R₁; R₃; R₄; R^(b); R^(c); and Prot^(NH) are as defined as above.

Further preferred compounds include compounds of general formula I, IA,IB, IC, IE, IF, IG, Ia, IAa, IBa, ICa, IEa, IFa, IGa, Ib, IAb, IBb, ICb,IEb, IFb, and IGb, wherein:

X is —NH—;

R₃ is hydrogen or a —OR^(b) group for compounds of formula I, IC, IE,IF, IG, Ia, ICa, IEa, IFa, IGa, Ib, ICb, IEb, IFb, or IGb; R₃ ishydrogen for compounds of formula IA, IAa, or IAb; and R₃ is a —OR^(b)group for compounds of formula IB, IBa or IBb; where R^(b) is asubstituted or unsubstituted C₁-C₆ alkyl. Particularly preferred R^(b)is selected from substituted or unsubstituted methyl, substituted orunsubstituted ethyl, substituted or unsubstituted n-propyl, substitutedor unsubstituted isopropyl, substituted or unsubstituted n-butyl,substituted or unsubstituted isobutyl, substituted or unsubstitutedsec-butyl and substituted or unsubstituted tert-butyl. More preferred R₃are hydrogen and methoxy, being hydrogen the most preferred R₃ group;and R₁; R₂; R₄; R^(a); R^(c); and Prot^(NH) are as defined as above.

Further preferred compounds include compounds of general formula I, IA,IB, IC, IE, IF, IG, Ia, IAa, IBa, ICa, IEa, IFa, IGa, Ib, IAb, IBb, ICb,IEb, IFb, and IGb, wherein:

X is —NH—;

R₄ is selected from —CH₂OH, —CH₂OC(═O)R^(c), —CH₂NH₂, and—CH₂NHProt^(NH) for compounds of formula I, IA, IB, IC, IF, IG, Ia, IAa,IBa, ICa, IFa, IGa, Ib, IAb, IBb, ICb, IFb, or IGb; and R₄ is selectedfrom —CH₂NH₂, and —CH₂NHProt^(NH) for compounds of formula IE, IEa orIEb; where R^(c) is a substituted or unsubstituted C₁-C₆ alkyl.Particularly preferred R^(c) is selected from substituted orunsubstituted methyl, substituted or unsubstituted ethyl, substituted orunsubstituted n-propyl, substituted or unsubstituted isopropyl,substituted or unsubstituted n-butyl, substituted or unsubstitutedisobutyl, substituted or unsubstituted sec-butyl, or substituted orunsubstituted tert-butyl. Most preferred R^(c) is methyl. More preferredR₄ is selected from CH₂OH and CH₂NH₂. More preferably, R₄ may be—CH₂NH₂. Most preferred R₄ is —CH₂OH; and R₁; R₂; R₃; R^(a); and R^(b)are as defined as above.

Further preferred compounds include compounds of general formula I, IA,IB, IC, IE, IF, IG, Ia, IAa, IBa, ICa, IEa, IFa, IGa, Ib, IAb, IBb, ICb,IEb, IFb, and IGb, wherein:

X is —NH—; R₁ is —OH;

R₂ is a —C(═O)R^(a) group for compounds of formula I, IA, IB, IC, IE,IF, Ia, IAa, IBa, ICa, IEa, IFa, Ib, IAb, IBb, ICb, IEb, or IFb; and R₂is acetyl for compounds of formula IG, IGa or IGb; where R^(a) is asubstituted or unsubstituted C₁-C₆ alkyl. Particularly preferred R^(a)is selected from substituted or unsubstituted methyl, substituted orunsubstituted ethyl, substituted or unsubstituted n-propyl, substitutedor unsubstituted isopropyl, substituted or unsubstituted n-butyl,substituted or unsubstituted isobutyl, substituted or unsubstitutedsec-butyl and substituted or unsubstituted tert-butyl. Most preferred R₂is acetyl;and R₃; R₄; R^(b); R^(c); and Prot^(NH) are as defined as above.

Further preferred compounds include compounds of general formula I, IA,IB, IC, IE, IF, IG, Ia, IAa, IBa, ICa, IEa, IFa, IGa, Ib, IAb, IBb, ICb,IEb, IFb, and IGb, wherein:

X is —NH—; R₁ is —OH;

R₃ is hydrogen or a —OR^(b) group for compounds of formula I, IC, IE,IF, IG, Ia, ICa, IEa, IFa, IGa, Ib, ICb, IEb, IFb, or IGb; R₃ ishydrogen for compounds of formula IA, IAa, or IAb; and R₃ is a —OR^(b)group for compounds of formula IB, IBa or IBb; where R^(b) is asubstituted or unsubstituted C₁-C₆ alkyl. Particularly preferred R^(b)is selected from substituted or unsubstituted methyl, substituted orunsubstituted ethyl, substituted or unsubstituted n-propyl, substitutedor unsubstituted isopropyl, substituted or unsubstituted n-butyl,substituted or unsubstituted isobutyl, substituted or unsubstitutedsec-butyl and substituted or unsubstituted tert-butyl. More preferred R₃are hydrogen and methoxy, being hydrogen the most preferred R₃ group;and R₂; R₄; R^(a); R^(c); and Prot^(NH) are as defined as above.

Further preferred compounds include compounds of general formula I, IA,IB, IC, IE, IF, IG, Ia, IAa, IBa, ICa, IEa, IFa, IGa, Ib, IAb, IBb, ICb,IEb, IFb, and IGb, wherein:

X is —NH—; R₁ is —OH;

R₄ is selected from —CH₂OH, —CH₂OC(═O)R^(c), —CH₂NH₂, and—CH₂NHProt^(NH) for compounds of formula I, IA, IB, IC, IF, IG, Ia, IAa,IBa, ICa, IFa, IGa, Ib, IAb, IBb, ICb, IFb, or IGb; and R₄ is selectedfrom —CH₂NH₂, and —CH₂NHProt^(NH) for compounds of formula IE, IEa orIEb; where is a substituted or unsubstituted C₁-C₆ alkyl. Particularlypreferred is selected from substituted or unsubstituted methyl,substituted or unsubstituted ethyl, substituted or unsubstitutedn-propyl, substituted or unsubstituted isopropyl, substituted orunsubstituted n-butyl, substituted or unsubstituted isobutyl,substituted or unsubstituted sec-butyl, and substituted or unsubstitutedtert-butyl. Most preferred is methyl. More preferred R₄ is selected fromCH₂OH and CH₂NH₂. More preferably, R₄ may be —CH₂NH₂. Most preferred R₄is —CH₂OH;and R₂; R₃; R^(a); and R^(b) are as defined as above.

Further preferred compounds include compounds of general formula I, IA,IB, IC, IE, IF, IG, Ia, IAa, IBa, ICa, IEa, IFa, IGa, Ib, IAb, IBb, ICb,IEb, IFb, and IGb, wherein:

X is —NH—;

R₂ is a —C(═O)R^(a) group for compounds of formula I, IA, IB, IC, IE,IF, Ia, IAa, IBa, ICa, IEa, IFa, Ib, IAb, IBb, ICb, IEb, or IFb; and R₂is acetyl for compounds of formula IG, IGa or IGb; where R^(a) is asubstituted or unsubstituted C₁-C₆ alkyl. Particularly preferred R^(a)is selected from substituted or unsubstituted methyl, substituted orunsubstituted ethyl, substituted or unsubstituted n-propyl, substitutedor unsubstituted isopropyl, substituted or unsubstituted n-butyl,substituted or unsubstituted isobutyl, substituted or unsubstitutedsec-butyl and substituted or unsubstituted tert-butyl. Most preferred R₂is acetyl;R₃ is hydrogen or a —OR^(b) group for compounds of formula I, IC, IE,IF, IG, Ia, ICa, IEa, IFa, IGa, Ib, ICb, IEb, IFb, or IGb; R₃ ishydrogen for compounds of formula IA, IAa, or IAb; and R₃ is a —OR^(b)group for compounds of formula IB, IBa or IBb; where R^(b) is asubstituted or unsubstituted C₁-C₆ alkyl. Particularly preferred R^(b)is selected from substituted or unsubstituted methyl, substituted orunsubstituted ethyl, substituted or unsubstituted n-propyl, substitutedor unsubstituted isopropyl, substituted or unsubstituted n-butyl,substituted or unsubstituted isobutyl, substituted or unsubstitutedsec-butyl and substituted or unsubstituted tert-butyl. More preferred R₃are hydrogen and methoxy, being hydrogen the most preferred R₃ group;and R₁; R₄; R^(c); and Prot^(NH) are as defined as above.

Further preferred compounds include compounds of general formula I, IA,IB, IC, IE, IF, IG, Ia, IAa, IBa, ICa, IEa, IFa, IGa, Ib, IAb, IBb, ICb,IEb, IFb, and IGb, wherein:

X is —NH—;

R₂ is a —C(═O)R^(a) group for compounds of formula I, IA, IB, IC, IE,IF, Ia, IAa, IBa, ICa, IEa, IFa, Ib, IAb, IBb, ICb, IEb, or IFb; and R₂is acetyl for compounds of formula IG, IGa or IGb; where R^(a) is asubstituted or unsubstituted C₁-C₆ alkyl. Particularly preferred R^(a)is selected from substituted or unsubstituted methyl, substituted orunsubstituted ethyl, substituted or unsubstituted n-propyl, substitutedor unsubstituted isopropyl, substituted or unsubstituted n-butyl,substituted or unsubstituted isobutyl, substituted or unsubstitutedsec-butyl and substituted or unsubstituted tert-butyl. Most preferred R₂is acetyl;R₄ is selected from —CH₂OH, —CH₂OC(═O)R^(c), —CH₂NH₂, and—CH₂NHProt^(NH) for compounds of formula I, IA, IB, IC, IF, IG, Ia, IAa,IBa, ICa, IFa, IGa, Ib, IAb, IBb, ICb, IFb, or IGb; and R₄ is selectedfrom —CH₂NH₂, and —CH₂NHProt^(NH) for compounds of formula IE, IEa orIEb; where R^(c) is a substituted or unsubstituted C₁-C₆ alkyl.Particularly preferred is selected from substituted or unsubstitutedmethyl, substituted or unsubstituted ethyl, substituted or unsubstitutedn-propyl, substituted or unsubstituted isopropyl, substituted orunsubstituted n-butyl, substituted or unsubstituted isobutyl,substituted or unsubstituted sec-butyl, and substituted or unsubstitutedtert-butyl. Most preferred R^(c) is methyl. More preferred R₄ isselected from CH₂OH and CH₂NH₂. More preferably, R₄ may be —CH₂NH₂. Mostpreferred R₄ is —CH₂OH;and R₁; R₃; and R^(b) are as defined as above.

Further preferred compounds include compounds of general formula I, IA,IB, IC, IE, IF, IG, Ia, IAa, IBa, ICa, IEa, IFa, IGa, Ib, IAb, IBb, ICb,IEb, IFb, and IGb, wherein:

X is —NH—;

R₃ is hydrogen or a —OR^(b) group for compounds of formula I, IC, IE,IF, IG, Ia, ICa, IEa, IFa, IGa, Ib, ICb, IEb, IFb, or IGb; R₃ ishydrogen for compounds of formula IA, IAa, or IAb; and R₃ is a —OR^(b)group for compounds of formula IB, IBa or IBb; where R^(b) is asubstituted or unsubstituted C₁-C₆ alkyl. Particularly preferred R^(b)is selected from substituted or unsubstituted methyl, substituted orunsubstituted ethyl, substituted or unsubstituted n-propyl, substitutedor unsubstituted isopropyl, substituted or unsubstituted n-butyl,substituted or unsubstituted isobutyl, substituted or unsubstitutedsec-butyl and substituted or unsubstituted tert-butyl. More preferred R₃are hydrogen and methoxy, being hydrogen the most preferred R₃ group;R₄ is selected from —CH₂OH, —CH₂OC(═O)R^(c), —CH₂NH₂, and—CH₂NHProt^(NH) for compounds of formula I, IA, IB, IC, IF, IG, Ia, IAa,IBa, ICa, IFa, IGa, Ib, IAb, IBb, ICb, IFb, or IGb; and R₄ is selectedfrom —CH₂NH₂, and —CH₂NHProt^(NH) for compounds of formula IE, IEa orIEb; whereR^(c) is a substituted or unsubstituted C₁-C₆ alkyl. Particularlypreferred R^(c) is selected from substituted or unsubstituted methyl,substituted or unsubstituted ethyl, substituted or unsubstitutedn-propyl, substituted or unsubstituted isopropyl, substituted orunsubstituted n-butyl, substituted or unsubstituted isobutyl,substituted or unsubstituted sec-butyl, and substituted or unsubstitutedtert-butyl. Most preferred is methyl. More preferred R₄ is selected fromCH₂OH and CH₂NH₂. More preferably, R₄ may be —CH₂NH₂. Most preferred R₄is —CH₂OH;and R₁; R₂; and R^(a); are as defined as above.

Further preferred compounds include compounds of general formula I, IA,IB, IC, IE, IF, IG, Ia, IAa, IBa, ICa, IEa, IFa, IGa, Ib, IAb, IBb, ICb,IEb, IFb, and IGb, wherein:

X is —NH—; R₁ is —OH;

R₂ is a —C(═O)R^(a) group for compounds of formula I, IA, IB, IC, IE,IF, Ia, IAa, IBa, ICa, IEa, IFa, Ib, IAb, IBb, ICb, IEb, or IFb; and R₂is acetyl for compounds of formula IG, IGa or IGb; where R^(a) is asubstituted or unsubstituted C₁-C₆ alkyl. Particularly preferred R^(a)is selected from substituted or unsubstituted methyl, substituted orunsubstituted ethyl, substituted or unsubstituted n-propyl, substitutedor unsubstituted isopropyl, substituted or unsubstituted n-butyl,substituted or unsubstituted isobutyl, substituted or unsubstitutedsec-butyl and substituted or unsubstituted tert-butyl. Most preferred R₂is acetyl;R₃ is hydrogen or a —OR^(b) group for compounds of formula I, IC, IE,IF, IG, Ia, ICa, IEa, IFa, IGa, Ib, ICb, IEb, IFb, or IGb; R₃ ishydrogen for compounds of formula IA, IAa, or IAb; and R₃ is a —OR^(b)group for compounds of formula IB, IBa or IBb; where R^(b) is asubstituted or unsubstituted C₁-C₆ alkyl. Particularly preferred R^(b)is selected from substituted or unsubstituted methyl, substituted orunsubstituted ethyl, substituted or unsubstituted n-propyl, substitutedor unsubstituted isopropyl, substituted or unsubstituted n-butyl,substituted or unsubstituted isobutyl, substituted or unsubstitutedsec-butyl and substituted or unsubstituted tert-butyl. More preferred R₃are hydrogen and methoxy, being hydrogen the most preferred R₃ group;and R₄; R^(c); and Prot^(NH) are as defined as above.

Further preferred compounds include compounds of general formula I, IA,IB, IC, IE, IF, IG, Ia, IAa, IBa, ICa, IEa, IFa, IGa, Ib, IAb, IBb, ICb,IEb, IFb, and IGb, wherein:

X is —NH—; R₁ is —OH;

R₂ is a —C(═O)R^(a) group for compounds of formula I, IA, IB, IC, IE,IF, Ia, IAa, IBa, ICa, IEa, IFa, Ib, IAb, IBb, ICb, IEb, or IFb; and R₂is acetyl for compounds of formula IG, IGa or IGb; where R^(a) is asubstituted or unsubstituted C₁-C₆ alkyl. Particularly preferred R^(a)is selected from substituted or unsubstituted methyl, substituted orunsubstituted ethyl, substituted or unsubstituted n-propyl, substitutedor unsubstituted isopropyl, substituted or unsubstituted n-butyl,substituted or unsubstituted isobutyl, substituted or unsubstitutedsec-butyl and substituted or unsubstituted tert-butyl. Most preferred R₂is acetyl;R₄ is selected from —CH₂OH, —CH₂OC(═O)R^(c), —CH₂NH₂, and—CH₂NHProt^(NH) for compounds of formula I, IA, IB, IC, IF, IG, Ia, IAa,IBa, ICa, IFa, IGa, Ib, IAb, IBb, ICb, IFb, or IGb; and R₄ is selectedfrom —CH₂NH₂, and —CH₂NHProt^(NH) for compounds of formula IE, IEa orIEb; where R^(c) is a substituted or unsubstituted C₁-C₆ alkyl.Particularly preferred is selected from substituted or unsubstitutedmethyl, substituted or unsubstituted ethyl, substituted or unsubstitutedn-propyl, substituted or unsubstituted isopropyl, substituted orunsubstituted n-butyl, substituted or unsubstituted isobutyl,substituted or unsubstituted sec-butyl, and substituted or unsubstitutedtert-butyl. Most preferred is methyl. More preferred R₄ is selected fromCH₂OH and CH₂NH₂. More preferably, R₄ may be —CH₂NH₂. Most preferred R₄is —CH₂OH;and R₃; and R^(b) are as defined as above.

Further preferred compounds include compounds of general formula I, IA,IB, IC, IE, IF, IG, Ia, IAa, IBa, ICa, IEa, IFa, IGa, Ib, IAb, IBb, ICb,IEb, IFb, and IGb, wherein:

X is —NH—;

R₂ is a —C(═O)R^(a) group for compounds of formula I, IA, IB, IC, IE,IF, Ia, IAa, IBa, ICa, IEa, IFa, Ib, IAb, IBb, ICb, IEb, or IFb; and R₂is acetyl for compounds of formula IG, IGa or IGb; where R^(a) is asubstituted or unsubstituted C₁-C₆ alkyl. Particularly preferred R^(a)is selected from substituted or unsubstituted methyl, substituted orunsubstituted ethyl, substituted or unsubstituted n-propyl, substitutedor unsubstituted isopropyl, substituted or unsubstituted n-butyl,substituted or unsubstituted isobutyl, substituted or unsubstitutedsec-butyl and substituted or unsubstituted tert-butyl. Most preferred R₂is acetyl;R₃ is hydrogen or a —OR^(b) group for compounds of formula I, IC, IE,IF, IG, Ia, ICa, IEa, IFa, IGa, Ib, ICb, IEb, IFb, or IGb; R₃ ishydrogen for compounds of formula IA, IAa, or IAb; and R₃ is a —OR^(b)group for compounds of formula IB, IBa or IBb; where R^(b) is asubstituted or unsubstituted C₁-C₆ alkyl. Particularly preferred R^(b)is selected from substituted or unsubstituted methyl, substituted orunsubstituted ethyl, substituted or unsubstituted n-propyl, substitutedor unsubstituted isopropyl, substituted or unsubstituted n-butyl,substituted or unsubstituted isobutyl, substituted or unsubstitutedsec-butyl and substituted or unsubstituted tert-butyl. More preferred R₃are hydrogen and methoxy, being hydrogen the most preferred R₃ group;R₄ is selected from —CH₂OH, —CH₂OC(═O)R^(c), —CH₂NH₂, and—CH₂NHProt^(NH) for compounds of formula I, IA, IB, IC, IF, IG, Ia, IAa,IBa, ICa, IFa, IGa, Ib, IAb, IBb, ICb, IFb, or IGb; and R₄ is selectedfrom —CH₂NH₂, and —CH₂NHProt^(NH) for compounds of formula IE, IEa orIEb; where is a substituted or unsubstituted C₁-C₆ alkyl. Particularlypreferred is selected from substituted or unsubstituted methyl,substituted or unsubstituted ethyl, substituted or unsubstitutedn-propyl, substituted or unsubstituted isopropyl, substituted orunsubstituted n-butyl, substituted or unsubstituted isobutyl,substituted or unsubstituted sec-butyl, and substituted or unsubstitutedtert-butyl. Most preferred is methyl. More preferred R₄ is selected fromCH₂OH and CH₂NH₂. More preferably, R₄ may be —CH₂NH₂. Most preferred R₄is —CH₂OH;and R₁ is as defined as above.

Further preferred compounds include compounds of general formula I, IA,IB, IC, IE, IF, IG, Ia, IAa, IBa, ICa, IEa, IFa, IGa, Ib, IAb, IBb, ICb,IEb, IFb, and IGb, wherein:

X is —NH—; R₁ is —OH;

R₂ is a —C(═O)R^(a) group for compounds of formula I, IA, IB, IC, IE,IF, Ia, IAa, IBa, ICa, IEa, IFa, Ib, IAb, IBb, ICb, IEb, or IFb; and R₂is acetyl for compounds of formula IG, IGa or IGb; where R^(a) is asubstituted or unsubstituted C₁-C₆ alkyl. Particularly preferred R^(a)is selected from substituted or unsubstituted methyl, substituted orunsubstituted ethyl, substituted or unsubstituted n-propyl, substitutedor unsubstituted isopropyl, substituted or unsubstituted n-butyl,substituted or unsubstituted isobutyl, substituted or unsubstitutedsec-butyl and substituted or unsubstituted tert-butyl. Most preferred R₂is acetyl;R₃ is hydrogen or a —OR^(b) group for compounds of formula I, IC, IE,IF, IG, Ia, ICa, IEa, IFa, IGa, Ib, ICb, IEb, IFb, or IGb; R₃ ishydrogen for compounds of formula IA, IAa, or IAb; and R₃ is a —OR^(b)group for compounds of formula IB, IBa or IBb; where R^(b) is asubstituted or unsubstituted C₁-C₆ alkyl. Particularly preferred R^(b)is selected from substituted or unsubstituted methyl, substituted orunsubstituted ethyl, substituted or unsubstituted n-propyl, substitutedor unsubstituted isopropyl, substituted or unsubstituted n-butyl,substituted or unsubstituted isobutyl, substituted or unsubstitutedsec-butyl and substituted or unsubstituted tert-butyl. More preferred R₃are hydrogen and methoxy, being hydrogen the most preferred R₃ group;R₄ is selected from —CH₂OH, —CH₂OC(═O)R^(c), —CH₂NH₂, and—CH₂NHProt^(NH) for compounds of formula I, IA, IB, IC, IF, IG, Ia, IAa,IBa, ICa, IFa, IGa, Ib, IAb, IBb, ICb, IFb, or IGb; and R₄ is selectedfrom —CH₂NH₂, and —CH₂NHProt^(NH) for compounds of formula IE, IEa orIEb; where R^(c) is a substituted or unsubstituted C₁-C₆ alkyl.Particularly preferred R^(c) is selected from substituted orunsubstituted methyl, substituted or unsubstituted ethyl, substituted orunsubstituted n-propyl, substituted or unsubstituted isopropyl,substituted or unsubstituted n-butyl, substituted or unsubstitutedisobutyl, substituted or unsubstituted sec-butyl, and substituted orunsubstituted tert-butyl. Most preferred is methyl. More preferred R₄ isselected from CH₂OH and CH₂NH₂. More preferably, R₄ may be —CH₂NH₂. Mostpreferred R₄ is —CH₂OH.

Further preferred compounds include compounds of general formula I, IA,IB, ID, IE, IF, IG, Ia, IAa, IBa, IDa, IEa, IFa, IGa, Ib, IAb, IBb, IDb,IEb, IFb, and IGb, wherein:

X is —O—; R₁ is —OH;

and R₂; R₃; R₄; R^(a); R^(b); R^(c); and Prot^(NH) are as defined asabove.

Further preferred compounds include compounds of general formula I, IA,IB, ID, IE, IF, IG, Ia, IAa, IBa, IDa, IEa, IFa, IGa, Ib, IAb, IBb, IDb,IEb, IFb, and IGb, wherein:

X is —O—;

R₂ is a —C(═O)R^(a) group for compounds of formula I, IA, IB, ID, IE,IF, Ia, IAa, IBa, IDa, IEa, IFa, Ib, IAb, IBb, IDb, IEb, or IFb; and R₂is acetyl for compounds of formula IG, IGa or IGb; where R^(a) is asubstituted or unsubstituted C₁-C₆ alkyl. Particularly preferred R^(a)is selected from substituted or unsubstituted methyl, substituted orunsubstituted ethyl, substituted or unsubstituted n-propyl, substitutedor unsubstituted isopropyl, substituted or unsubstituted n-butyl,substituted or unsubstituted isobutyl, substituted or unsubstitutedsec-butyl and substituted or unsubstituted tert-butyl. Most preferred R₂is acetyl;

and R₁; R₃; R₄; R^(b); R^(c); and Prot^(NH) are as defined as above.

Further preferred compounds include compounds of general formula I, IA,IB, ID, IE, IF, IG, Ia, IAa, IBa, IDa, IEa, IFa, IGa, Ib, IAb, IBb, IDb,IEb, IFb, and IGb, wherein:

X is —O—;

R₃ is hydrogen or a —OR^(b) group for compounds of formula I, ID, IE,IF, IG, Ia, IDa, IEa, IFa, IGa, Ib, IDb, IEb, IFb, or IGb; R₃ ishydrogen for compounds of formula IA, IAa, or IAb; and R₃ is a —OR^(b)group for compounds of formula IB, IBa or IBb; where R^(b) is asubstituted or unsubstituted C₁-C₆ alkyl. Particularly preferred R^(b)is selected from substituted or unsubstituted methyl, substituted orunsubstituted ethyl, substituted or unsubstituted n-propyl, substitutedor unsubstituted isopropyl, substituted or unsubstituted n-butyl,substituted or unsubstituted isobutyl, substituted or unsubstitutedsec-butyl and substituted or unsubstituted tert-butyl. More preferred R₃is hydrogen and methoxy, being hydrogen the most preferred R₃ group; andR₁; R₂; R₄; R^(a); R^(c); and Prot^(NH) are as defined as above.

Further preferred compounds include compounds of general formula I, IA,IB, ID, IE, IF, IG, Ia, IAa, IBa, IDa, IEa, IFa, IGa, Ib, IAb, IBb, IDb,IEb, IFb, and IGb, wherein:

X is —O—;

R₄ is selected from —CH₂OH, —CH₂OC(═O)R^(c), —CH₂NH₂, and—CH₂NHProt^(NH) for compounds of formula I, IA, IB, ID, IF, IG, Ia, IAa,IBa, IDa, IFa, IGa, Ib, IAb, IBb, IDb, IFb, or IGb; and R₄ is selectedfrom —CH₂NH₂, and —CH₂NHProt^(NH) for compounds of formula IE, IEa orIEb; where R^(c) is a substituted or unsubstituted C₁-C₆ alkyl.Particularly preferred is selected from substituted or unsubstitutedmethyl, substituted or unsubstituted ethyl, substituted or unsubstitutedn-propyl, substituted or unsubstituted isopropyl, substituted orunsubstituted n-butyl, substituted or unsubstituted isobutyl,substituted or unsubstituted sec-butyl, and substituted or unsubstitutedtert-butyl. Most preferred is methyl. More preferred R₄ is selected from—CH₂OH and CH₂NH₂. More preferably, R₄ may be —CH₂NH₂. Most preferred R₄is —CH₂OH;and R₁; R₂; R₃; R^(a); and R^(b) are as defined as above.

Further preferred compounds include compounds of general formula I, IA,IB, ID, IE, IF, IG, Ia, IAa, IBa, IDa, IEa, IFa, IGa, Ib, IAb, IBb, IDb,IEb, IFb, and IGb, wherein:

X is —O—; R₁ is —OH;

R₂ is a —C(═O)R^(a) group for compounds of formula I, IA, IB, ID, IE,IF, Ia, IAa, IBa, IDa, IEa, IFa, Ib, IAb, IBb, IDb, IEb, or IFb; and R₂is acetyl for compounds of formula IG, IGa or IGb; where R^(a) is asubstituted or unsubstituted C₁-C₆ alkyl. Particularly preferred R^(a)is selected from substituted or unsubstituted methyl, substituted orunsubstituted ethyl, substituted or unsubstituted n-propyl, substitutedor unsubstituted isopropyl, substituted or unsubstituted n-butyl,substituted or unsubstituted isobutyl, substituted or unsubstitutedsec-butyl and substituted or unsubstituted tert-butyl. Most preferred R₂is acetyl;and R₃; R₄; R^(b); R^(c); and Prot^(NH) are as defined as above.

Further preferred compounds include compounds of general formula I, IA,IB, ID, IE, IF, IG, Ia, IAa, IBa, IDa, IEa, IFa, IGa, Ib, IAb, IBb, IDb,IEb, IFb, and IGb, wherein:

X is —O—; R₁ is —OH;

R₃ is hydrogen or a —OR^(b) group for compounds of formula I, ID, IE,IF, IG, Ia, IDa, IEa, IFa, IGa, Ib, IDb, IEb, IFb, or IGb; R₃ ishydrogen for compounds of formula IA, IAa, or IAb; and R₃ is a —OR^(b)group for compounds of formula IB, IBa or IBb; where R^(b) is asubstituted or unsubstituted C₁-C₆ alkyl. Particularly preferred R^(b)is selected from substituted or unsubstituted methyl, substituted orunsubstituted ethyl, substituted or unsubstituted n-propyl, substitutedor unsubstituted isopropyl, substituted or unsubstituted n-butyl,substituted or unsubstituted isobutyl, substituted or unsubstitutedsec-butyl and substituted or unsubstituted tert-butyl. More preferred R₃are hydrogen and methoxy, being hydrogen the most preferred R₃ group;and R₂; R₄; R^(a); R^(c); and Prot^(NH) are as defined as above.

Further preferred compounds include compounds of general formula I, IA,IB, ID, IE, IF, IG, Ia, IAa, IBa, IDa, IEa, IFa, IGa, Ib, IAb, IBb, IDb,IEb, IFb, and IGb, wherein:

X is —O—; R₁ is —OH;

R₄ is selected from —CH₂OH, —CH₂OC(═O)R^(c), —CH₂NH₂, and—CH₂NHProt^(NH) for compounds of formula I, IA, IB, ID, IF, IG, Ia, IAa,IBa, IDa, IFa, IGa, Ib, IAb, IBb, IDb, IFb, or IGb; and R₄ is selectedfrom —CH₂NH₂, and —CH₂NHProt^(NH) for compounds of formula IE, IEa orIEb; where R^(c) is a substituted or unsubstituted C₁-C₆ alkyl.Particularly preferred R^(c) is selected from substituted orunsubstituted methyl, substituted or unsubstituted ethyl, substituted orunsubstituted n-propyl, substituted or unsubstituted isopropyl,substituted or unsubstituted n-butyl, substituted or unsubstitutedisobutyl, substituted or unsubstituted sec-butyl, and substituted orunsubstituted tert-butyl. Most preferred is methyl. More preferred R₄ isselected from —CH₂OH and CH₂NH₂. More preferably, R₄ may be —CH₂NH₂.Most preferred R₄ is —CH₂OH;and R₂; R₃; R^(a); and R^(b) are as defined as above.

Further preferred compounds include compounds of general formula I, IA,IB, ID, IE, IF, IG, Ia, IAa, IBa, IDa, IEa, IFa, IGa, Ib, IAb, IBb, IDb,IEb, IFb, and IGb, wherein:

X is —O—;

R₂ is a —C(═O)R^(a) group for compounds of formula I, IA, IB, ID, IE,IF, Ia, IAa, IBa, IDa, IEa, IFa, Ib, IAb, IBb, IDb, IEb, or IFb; and R₂is acetyl for compounds of formula IG, IGa or IGb; where R^(a) is asubstituted or unsubstituted C₁-C₆ alkyl. Particularly preferred R^(a)is selected from substituted or unsubstituted methyl, substituted orunsubstituted ethyl, substituted or unsubstituted n-propyl, substitutedor unsubstituted isopropyl, substituted or unsubstituted n-butyl,substituted or unsubstituted isobutyl, substituted or unsubstitutedsec-butyl and substituted or unsubstituted tert-butyl. Most preferred R₂is acetyl;R₃ is hydrogen or a —OR^(b) group for compounds of formula I, ID, IE,IF, IG, Ia, IDa, IEa, IFa, IGa, Ib, IDb, IEb, IFb, or IGb; R₃ ishydrogen for compounds of formula IA, IAa, or IAb; and R₃ is a —OR^(b)group for compounds of formula IB, IBa or IBb; where R^(b) is asubstituted or unsubstituted C₁-C₆ alkyl. Particularly preferred R^(b)is selected from substituted or unsubstituted methyl, substituted orunsubstituted ethyl, substituted or unsubstituted n-propyl, substitutedor unsubstituted isopropyl, substituted or unsubstituted n-butyl,substituted or unsubstituted isobutyl, substituted or unsubstitutedsec-butyl and substituted or unsubstituted tert-butyl. More preferred R₃are hydrogen and methoxy, being hydrogen the most preferred R₃ group;and R₁; R₄; R^(c); and Prot^(NH) are as defined as above.

Further preferred compounds include compounds of general formula I, IA,IB, ID, IE, IF, IG, Ia, IAa, IBa, IDa, IEa, IFa, IGa, Ib, IAb, IBb, IDb,IEb, IFb, and IGb, wherein:

X is —O—;

R₂ is a —C(═O)R^(a) group for compounds of formula I, IA, IB, ID, IE,IF, Ia, IAa, IBa, IDa, IEa, IFa, Ib, IAb, IBb, IDb, IEb, or IFb; and R₂is acetyl for compounds of formula IG, IGa or IGb; where R^(a) is asubstituted or unsubstituted C₁-C₆ alkyl. Particularly preferred R^(a)is selected from substituted or unsubstituted methyl, substituted orunsubstituted ethyl, substituted or unsubstituted n-propyl, substitutedor unsubstituted isopropyl, substituted or unsubstituted n-butyl,substituted or unsubstituted isobutyl, substituted or unsubstitutedsec-butyl and substituted or unsubstituted tert-butyl. Most preferred R₂is acetyl;R₄ is selected from —CH₂OH, —CH₂OC(═O)R^(c), —CH₂NH₂, and—CH₂NHProt^(NH) for compounds of formula I, IA, IB, ID, IF, IG, Ia, IAa,IBa, IDa, IFa, IGa, Ib, IAb, IBb, IDb, IFb, or IGb; and R₄ is selectedfrom —CH₂NH₂, and —CH₂NHProt^(NH) for compounds of formula IE, IEa orIEb; where R^(c) is a substituted or unsubstituted C₁-C₆ alkyl.Particularly preferred is selected from substituted or unsubstitutedmethyl, substituted or unsubstituted ethyl, substituted or unsubstitutedn-propyl, substituted or unsubstituted isopropyl, substituted orunsubstituted n-butyl, substituted or unsubstituted isobutyl,substituted or unsubstituted sec-butyl, and substituted or unsubstitutedtert-butyl. Most preferred is methyl. More preferred R₄ is selected from—CH₂OH and —CH₂NH₂. More preferably, R₄ may be —CH₂NH₂. Most preferredR₄ is —CH₂OH;and R₁; R₃; and R^(b) are as defined as above.

Further preferred compounds include compounds of general formula I, IA,IB, ID, IE, IF, IG, Ia, IAa, IBa, IDa, IEa, IFa, IGa, Ib, IAb, IBb, IDb,IEb, IFb, and IGb, wherein:

X is —O—;

R₃ is hydrogen or a —OR^(b) group for compounds of formula I, ID, IE,IF, IG, Ia, IDa, IEa, IFa, IGa, Ib, IDb, IEb, IFb, or IGb; R₃ ishydrogen for compounds of formula IA, IAa, or IAb; and R₃ is a —OR^(b)group for compounds of formula IB, IBa or IBb; where R^(b) is asubstituted or unsubstituted C₁-C₆ alkyl. Particularly preferred R^(b)is selected from substituted or unsubstituted methyl, substituted orunsubstituted ethyl, substituted or unsubstituted n-propyl, substitutedor unsubstituted isopropyl, substituted or unsubstituted n-butyl,substituted or unsubstituted isobutyl, substituted or unsubstitutedsec-butyl and substituted or unsubstituted tert-butyl. More preferred R₃are hydrogen and methoxy, being hydrogen the most preferred R₃ group;R₄ is selected from —CH₂OH, —CH₂OC(═O)R^(c), —CH₂NH₂, and—CH₂NHProt^(NH) for compounds of formula I, IA, IB, ID, IF, IG, Ia, IAa,IBa, IDa, IFa, IGa, Ib, IAb, IBb, IDb, IFb, or IGb; and R₄ is selectedfrom —CH₂NH₂, and —CH₂NHProt^(NH) for compounds of formula IE, IEa orIEb; where R^(c) is a substituted or unsubstituted C₁-C₆ alkyl.Particularly preferred is selected from substituted or unsubstitutedmethyl, substituted or unsubstituted ethyl, substituted or unsubstitutedn-propyl, substituted or unsubstituted isopropyl, substituted orunsubstituted n-butyl, substituted or unsubstituted isobutyl,substituted or unsubstituted sec-butyl, and substituted or unsubstitutedtert-butyl. Most preferred R^(c) is methyl. More preferred R₄ isselected from —CH₂OH and —CH₂NH₂. More preferably, R₄ may be —CH₂NH₂.Most preferred R₄ is —CH₂OH;and R₁; R₂; and R^(a); are as defined as above.

Further preferred compounds include compounds of general formula I, IA,IB, ID, IE, IF, IG, Ia, IAa, IBa, IDa, IEa, IFa, IGa, Ib, IAb, IBb, IDb,IEb, IFb, and IGb, wherein:

X is —O—; R₁ is —OH;

R₂ is a —C(═O)R^(a) group for compounds of formula I, IA, IB, ID, IE,IF, Ia, IAa, IBa, IDa, IEa, IFa, Ib, IAb, IBb, IDb, IEb, or IFb; and R₂is acetyl for compounds of formula IG, IGa or IGb; where R^(a) is asubstituted or unsubstituted C₁-C₆ alkyl. Particularly preferred R^(a)is selected from substituted or unsubstituted methyl, substituted orunsubstituted ethyl, substituted or unsubstituted n-propyl, substitutedor unsubstituted isopropyl, substituted or unsubstituted n-butyl,substituted or unsubstituted isobutyl, substituted or unsubstitutedsec-butyl and substituted or unsubstituted tert-butyl. Most preferred R₂is acetyl;R₃ is hydrogen or a —OR^(b) group for compounds of formula I, ID, IE,IF, IG, Ia, IDa, IEa, IFa, IGa, Ib, IDb, IEb, IFb, or IGb; R₃ ishydrogen for compounds of formula IA, IAa, or IAb; and R₃ is a —OR^(b)group for compounds of formula IB, IBa or IBb; where R^(b) is asubstituted or unsubstituted C₁-C₆ alkyl. Particularly preferred R^(b)is selected from substituted or unsubstituted methyl, substituted orunsubstituted ethyl, substituted or unsubstituted n-propyl, substitutedor unsubstituted isopropyl, substituted or unsubstituted n-butyl,substituted or unsubstituted isobutyl, substituted or unsubstitutedsec-butyl and substituted or unsubstituted tert-butyl. More preferred R₃are hydrogen and methoxy, being hydrogen the most preferred R₃ group;and R₄; R^(c); and Prot^(NH) are as defined as above.

Further preferred compounds include compounds of general formula I, IA,IB, ID, IE, IF, IG, Ia, IAa, IBa, IDa, IEa, IFa, IGa, Ib, IAb, IBb, IDb,IEb, IFb, and IGb, wherein:

X is —O—; R₁ is —OH;

R₂ is a —C(═O)R^(a) group for compounds of formula I, IA, IB, ID, IE,IF, Ia, IAa, IBa, IDa, IEa, IFa, Ib, IAb, IBb, IDb, IEb, or IFb; and R₂is acetyl for compounds of formula IG, IGa or IGb; where R^(a) is asubstituted or unsubstituted C₁-C₆ alkyl. Particularly preferred R^(a)is selected from substituted or unsubstituted methyl, substituted orunsubstituted ethyl, substituted or unsubstituted n-propyl, substitutedor unsubstituted isopropyl, substituted or unsubstituted n-butyl,substituted or unsubstituted isobutyl, substituted or unsubstitutedsec-butyl and substituted or unsubstituted tert-butyl. Most preferred R₂is acetyl;R₄ is selected from —CH₂OH, —CH₂OC(═O)R^(c), —CH₂NH₂, and—CH₂NHProt^(NH) for compounds of formula I, IA, IB, ID, IF, IG, Ia, IAa,IBa, IDa, IFa, IGa, Ib, IAb, IBb, IDb, IFb, or IGb; and R₄ is selectedfrom —CH₂NH₂, and —CH₂NHProt^(NH) for compounds of formula IE, IEa orIEb; where R^(c) is a substituted or unsubstituted C₁-C₆ alkyl.Particularly preferred is selected from substituted or unsubstitutedmethyl, substituted or unsubstituted ethyl, substituted or unsubstitutedn-propyl, substituted or unsubstituted isopropyl, substituted orunsubstituted n-butyl, substituted or unsubstituted isobutyl,substituted or unsubstituted sec-butyl, and substituted or unsubstitutedtert-butyl. Most preferred is methyl. More preferred R₄ is selected from—CH₂OH and —CH₂NH₂. More preferably, R₄ may be —CH₂NH₂. Most preferredR₄ is —CH₂OH;

and R₃; and R^(b) are as defined as above.

Further preferred compounds include compounds of general formula I, IA,IB, ID, IE, IF, IG, Ia, IAa, IBa, IDa, IEa, IFa, IGa, Ib, IAb, IBb, IDb,IEb, IFb, and IGb, wherein:

X is —O—;

R₂ is a —C(═O)R^(a) group for compounds of formula I, IA, IB, ID, IE,IF, Ia, IAa, IBa, IDa, IEa, IFa, Ib, IAb, IBb, IDb, IEb, or IFb; and R₂is acetyl for compounds of formula IG, IGa or IGb; where R^(a) is asubstituted or unsubstituted C₁-C₆ alkyl. Particularly preferred R^(a)is selected from substituted or unsubstituted methyl, substituted orunsubstituted ethyl, substituted or unsubstituted n-propyl, substitutedor unsubstituted isopropyl, substituted or unsubstituted n-butyl,substituted or unsubstituted isobutyl, substituted or unsubstitutedsec-butyl and substituted or unsubstituted tert-butyl. Most preferred R₂is acetyl;R₃ is hydrogen or a —OR^(b) group for compounds of formula I, ID, IE,IF, IG, Ia, IDa, IEa, IFa, IGa, Ib, IDb, IEb, IFb, or IGb; R₃ ishydrogen for compounds of formula IA, IAa, or IAb; and R₃ is a —OR^(b)group for compounds of formula IB, IBa or IBb; where R^(b) is asubstituted or unsubstituted C₁-C₆ alkyl. Particularly preferred R^(b)is selected from substituted or unsubstituted methyl, substituted orunsubstituted ethyl, substituted or unsubstituted n-propyl, substitutedor unsubstituted isopropyl, substituted or unsubstituted n-butyl,substituted or unsubstituted isobutyl, substituted or unsubstitutedsec-butyl and substituted or unsubstituted tert-butyl. More preferred R₃are hydrogen and methoxy, being hydrogen the most preferred R₃ group;R₄ is selected from —CH₂OH, —CH₂OC(═O)R^(c), —CH₂NH₂, and—CH₂NHProt^(NH) for compounds of formula I, IA, IB, ID, IF, IG, Ia, IAa,IBa, IDa, IFa, IGa, Ib, IAb, IBb, IDb, IFb, or IGb; and R₄ is selectedfrom —CH₂NH₂, and —CH₂NHProt^(NH) for compounds of formula IE, IEa orIEb; where R^(c) is a substituted or unsubstituted C₁-C₆ alkyl.Particularly preferred R^(c) is selected from substituted orunsubstituted methyl, substituted or unsubstituted ethyl, substituted orunsubstituted n-propyl, substituted or unsubstituted isopropyl,substituted or unsubstituted n-butyl, substituted or unsubstitutedisobutyl, substituted or unsubstituted sec-butyl, and substituted orunsubstituted tert-butyl. Most preferred is methyl. More preferred R₄ isselected from —CH₂OH and —CH₂NH₂. More preferably, R₄ may be —CH₂NH₂.Most preferred R₄ is —CH₂OH;

and R₁ is as defined as above.

Further preferred compounds include compounds of general formula I, IA,IB, ID, IE, IF, IG, Ia, IAa, IBa, IDa, IEa, IFa, IGa, Ib, IAb, IBb, IDb,IEb, IFb, and IGb, wherein:

X is —O—; R₁ is —OH;

R₂ is a —C(═O)R^(a) group for compounds of formula I, IA, IB, ID, IE,IF, Ia, IAa, IBa, IDa, IEa, IFa, Ib, IAb, IBb, IDb, IEb, or IFb; and R₂is acetyl for compounds of formula IG, IGa or IGb; where R^(a) is asubstituted or unsubstituted C₁-C₆ alkyl. Particularly preferred R^(a)is selected from substituted or unsubstituted methyl, substituted orunsubstituted ethyl, substituted or unsubstituted n-propyl, substitutedor unsubstituted isopropyl, substituted or unsubstituted n-butyl,substituted or unsubstituted isobutyl, substituted or unsubstitutedsec-butyl and substituted or unsubstituted tert-butyl. Most preferred R₂is acetyl;R₃ is hydrogen or a —OR^(b) group for compounds of formula I, ID, IE,IF, IG, Ia, IDa, IEa, IFa, IGa, Ib, IDb, IEb, IFb, or IGb; R₃ ishydrogen for compounds of formula IA, IAa, or IAb; and R₃ is a —OR^(b)group for compounds of formula IB, IBa or IBb; where R^(b) is asubstituted or unsubstituted C₁-C₆ alkyl. Particularly preferred R^(b)is selected from substituted or unsubstituted methyl, substituted orunsubstituted ethyl, substituted or unsubstituted n-propyl, substitutedor unsubstituted isopropyl, substituted or unsubstituted n-butyl,substituted or unsubstituted isobutyl, substituted or unsubstitutedsec-butyl and substituted or unsubstituted tert-butyl. More preferred R₃are hydrogen and methoxy, being hydrogen the most preferred R₃ group;R₄ is selected from —CH₂OH, —CH₂OC(═O)R^(c), —CH₂NH₂, and—CH₂NHProt^(NH) for compounds of formula I, IA, IB, ID, IF, IG, Ia, IAa,IBa, IDa, IFa, IGa, Ib, IAb, IBb, IDb, IFb, or IGb; and R₄ is selectedfrom —CH₂NH₂, and —CH₂NHProt^(NH) for compounds of formula IE, IEa orIEb; where R^(c) is a substituted or unsubstituted C₁-C₆ alkyl.Particularly preferred is selected from substituted or unsubstitutedmethyl, substituted or unsubstituted ethyl, substituted or unsubstitutedn-propyl, substituted or unsubstituted isopropyl, substituted orunsubstituted n-butyl, substituted or unsubstituted isobutyl,substituted or unsubstituted sec-butyl, and substituted or unsubstitutedtert-butyl. Most preferred R^(c) is methyl. More preferred R₄ isselected from —CH₂OH and —CH₂NH₂. More preferably, R₄ may be —CH₂NH₂.Most preferred R₄ is —CH₂OH.

Further preferred compounds include compounds of general formula Ic,IAc, IBc, IDc, and IGc wherein:

R₁ is —OH;

and R₂; R₃; R^(a) and R^(b) are as defined as above.

Further preferred compounds include compounds of general formula Ic,IAc, IBc, IDc, IFc, and IGc, wherein:

R₂ is a —C(═O)R^(a) group for compounds of formula Ic, IAc, IBc, IDc, orIFc; and R₂ is acetyl for compounds of formula IGc; where R^(a) is asubstituted or unsubstituted C₁-C₆ alkyl. Particularly preferred R^(a)is selected from substituted or unsubstituted methyl, substituted orunsubstituted ethyl, substituted or unsubstituted n-propyl, substitutedor unsubstituted isopropyl, substituted or unsubstituted n-butyl,substituted or unsubstituted isobutyl, substituted or unsubstitutedsec-butyl and substituted or unsubstituted tert-butyl. Most preferred R₂is acetyl;and R₁; R₃; R^(b) are as defined as above.

Further preferred compounds include compounds of general formula Ic,IAc, IBc, IDc, IFc, and IGc, wherein:

R₃ is hydrogen or a —OR^(b) group for compounds of formula Ic, IDc, IFc,or IGc; R₃ is hydrogen for compounds of formula IAc; and R₃ is a —OR^(b)group for compounds of formula IBc; where R^(b) is a substituted orunsubstituted C₁-C₆ alkyl. Particularly preferred R^(b) is selected fromsubstituted or unsubstituted methyl, substituted or unsubstituted ethyl,substituted or unsubstituted n-propyl, substituted or unsubstitutedisopropyl, substituted or unsubstituted n-butyl, substituted orunsubstituted isobutyl, substituted or unsubstituted sec-butyl andsubstituted or unsubstituted tert-butyl. More preferred R₃ are hydrogenand methoxy, being hydrogen the most preferred R₃ group;and R₁; R₂; and IV are as defined as above.

Further preferred compounds include compounds of general formula Ic,IAc, IBc, IDc, IFc, and IGc, wherein:

R₁ is —OH;

R₂ is a —C(═O)R^(a) group for compounds of formula Ic, IAc, IBc, IDc, orIFc; and R₂ is acetyl for compounds of formula IGc; where R^(a) is asubstituted or unsubstituted C₁-C₆ alkyl. Particularly preferred R^(a)is selected from substituted or unsubstituted methyl, substituted orunsubstituted ethyl, substituted or unsubstituted n-propyl, substitutedor unsubstituted isopropyl, substituted or unsubstituted n-butyl,substituted or unsubstituted isobutyl, substituted or unsubstitutedsec-butyl and substituted or unsubstituted tert-butyl. Most preferred R₂is acetyl; and R₃; and R^(b) are as defined as above.

Further preferred compounds include compounds of general formula Ic,IAc, IBc, IDc, IFc, and IGc, wherein:

R₁ is —OH;

R₃ is hydrogen or a —OR^(b) group for compounds of formula Ic, IDc, IFc,or IGc; R₃ is hydrogen for compounds of formula IAc; and R₃ is a —OR^(b)group for compounds of formula IBc; where R^(b) is a substituted orunsubstituted C₁-C₆ alkyl. Particularly preferred R^(b) is selected fromsubstituted or unsubstituted methyl, substituted or unsubstituted ethyl,substituted or unsubstituted n-propyl, substituted or unsubstitutedisopropyl, substituted or unsubstituted n-butyl, substituted orunsubstituted isobutyl, substituted or unsubstituted sec-butyl andsubstituted or unsubstituted tert-butyl. More preferred R₃ are hydrogenand methoxy, being hydrogen the most preferred R₃ group;and R₂; and IV are as defined as above.

Further preferred compounds include compounds of general formula Ic,IAc, IBc, IDc, IFc, and IGc, wherein:

R₂ is a —C(═O)R^(a) group for compounds of formula Ic, IAc, IBc, IDc, orIFc; and R₂ is acetyl for compounds of formula IGc; where R^(a) is asubstituted or unsubstituted C₁-C₆ alkyl. Particularly preferred R^(a)is selected from substituted or unsubstituted methyl, substituted orunsubstituted ethyl, substituted or unsubstituted n-propyl, substitutedor unsubstituted isopropyl, substituted or unsubstituted n-butyl,substituted or unsubstituted isobutyl, substituted or unsubstitutedsec-butyl and substituted or unsubstituted tert-butyl. Most preferred R₂is acetyl;R₃ is hydrogen or a —OR^(b) group for compounds of formula Ic, IDc, IFc,or IGc; R₃ is hydrogen for compounds of formula IAc; and R₃ is a —OR^(b)group for compounds of formula IBc; where R^(b) is a substituted orunsubstituted C₁-C₆ alkyl. Particularly preferred R^(b) is selected fromsubstituted or unsubstituted methyl, substituted or unsubstituted ethyl,substituted or unsubstituted n-propyl, substituted or unsubstitutedisopropyl, substituted or unsubstituted n-butyl, substituted orunsubstituted isobutyl, substituted or unsubstituted sec-butyl andsubstituted or unsubstituted tert-butyl. More preferred R₃ are hydrogenand methoxy, being hydrogen the most preferred R₃ group;and R₁ is as defined as above.

Further preferred compounds include compounds of general formula Ic,IAc, IBc, IDc, IFc, and IGc, wherein:

R₁ is —OH;

R₂ is a —C(═O)R^(a) group for compounds of formula Ic, IAc, IBc, IDc, orIFc; and R₂ is acetyl for compounds of formula IGc; where R^(a) is asubstituted or unsubstituted C₁-C₆ alkyl. Particularly preferred R^(a)is selected from substituted or unsubstituted methyl, substituted orunsubstituted ethyl, substituted or unsubstituted n-propyl, substitutedor unsubstituted isopropyl, substituted or unsubstituted n-butyl,substituted or unsubstituted isobutyl, substituted or unsubstitutedsec-butyl and substituted or unsubstituted tert-butyl. Most preferred R₂is acetyl;R₃ is hydrogen or a —OR^(b) group for compounds of formula Ic, IDc, IFc,or IGc; R₃ is hydrogen for compounds of formula IAc; and R₃ is a —OR^(b)group for compounds of formula IBc; where R^(b) is a substituted orunsubstituted C₁-C₆ alkyl. Particularly preferred R^(b) is selected fromsubstituted or unsubstituted methyl, substituted or unsubstituted ethyl,substituted or unsubstituted n-propyl, substituted or unsubstitutedisopropyl, substituted or unsubstituted n-butyl, substituted orunsubstituted isobutyl, substituted or unsubstituted sec-butyl andsubstituted or unsubstituted tert-butyl. More preferred R₃ are hydrogenand methoxy, being hydrogen the most preferred R₃ group.

The following preferred substituents (where allowed by possiblesubstituent groups) apply to compounds of formula I, IA, IB, IC, ID, IE,IF, IG, Ia, IAa, IBa, ICa, IDa, IEa, IFa, IGa, Ib, IAb, IBb, ICb, IDb,IEb, IFb, IGb, Ic, IAc, IBc, IDc, IFc, and IGc:

In compounds of the present invention, particularly preferred R₁ is —OH.

In compounds of the present invention, particularly preferred R₂ is a—C(═O)R^(a) group where R^(a) is a substituted or unsubstituted C₁-C₆alkyl. Particularly preferred R^(a) is selected from substituted orunsubstituted methyl, substituted or unsubstituted ethyl, substituted orunsubstituted n-propyl, substituted or unsubstituted isopropyl,substituted or unsubstituted n-butyl, substituted or unsubstitutedisobutyl, substituted or unsubstituted sec-butyl and substituted orunsubstituted tert-butyl. Most preferred R₂ is acetyl.

In compounds of the present invention, particularly preferred R₃ ishydrogen or a —OR^(b) group where R^(b) is a substituted orunsubstituted C₁-C₆ alkyl. Particularly preferred R^(b) is selected fromsubstituted or unsubstituted methyl, substituted or unsubstituted ethyl,substituted or unsubstituted n-propyl, substituted or unsubstitutedisopropyl, substituted or unsubstituted n-butyl, substituted orunsubstituted isobutyl, substituted or unsubstituted sec-butyl andsubstituted or unsubstituted tert-butyl. More preferred R₃ are hydrogenand methoxy, being hydrogen the most preferred R₃ group.

In compounds of the present invention, particularly preferred R₄ isselected from H, —CH₂OH, —CH₂OC(═O)R^(c), —CH₂NH₂, and —CH₂NHProt^(NH)where is a substituted or unsubstituted C₁-C₆ alkyl. Particularlypreferred is selected from substituted or unsubstituted methyl,substituted or unsubstituted ethyl, substituted or unsubstitutedn-propyl, substituted or unsubstituted isopropyl, substituted orunsubstituted n-butyl, substituted or unsubstituted isobutyl,substituted or unsubstituted sec-butyl, and substituted or unsubstitutedtert-butyl. Most preferred R^(c) is methyl. More preferred R₄ isselected from H, CH₂OH and CH₂NH₂. Most preferred R₄ is —CH₂OH.

In compounds of general formula I, IA, IB, IC, ID, IE, IF, IG, Ia, IAa,IBa, ICa, IDa, IEa, IFa, IGa, Ib, IAb, IBb, ICb, IDb, IEb, IFb, and IGbparticularly preferred R₄ is selected from —CH₂OH, —CH₂OC(═O)R^(c),—CH₂NH₂, and —CH₂NHProt^(NH) for compounds of formula I, IA, IB, IC, ID,IF, IG, Ia, IAa, IBa, ICa, IDa, IFa, IGa, Ib, IAb, IBb, ICb, IDb, IFb,or IGb; and R₄ is selected from —CH₂NH₂, and —CH₂NHProt^(NH) forcompounds of formula IE, IEa or IEb; where R^(c) is a substituted orunsubstituted C₁-C₆ alkyl. Particularly preferred R^(c) is a substitutedor unsubstituted methyl, substituted or unsubstituted ethyl, substitutedor unsubstituted n-propyl, substituted or unsubstituted isopropyl,substituted or unsubstituted n-butyl, substituted or unsubstitutedisobutyl, substituted or unsubstituted sec-butyl, and substituted orunsubstituted tert-butyl. Most preferred R^(c) is methyl. More preferredR₄ is selected from CH₂OH and CH₂NH₂. Most preferred R₄ is —CH₂OH.

Being particularly preferred compounds of formula Ia, IAa, IBa, ICa,IDa, IFa, IGa when R₄ is —CH₂OH or —CH₂OC(═O)R^(c) and compounds offormula Ib, IAb, IBb, ICb, IDb, IEb, IFb, IGb when R₄ is —CH₂NH₂ or—CH₂NHProt^(NH).

In compounds of the present invention, particularly preferred X is —NH—.

Alternatively, in compounds of the present invention, particularlypreferred X is —O—.

Preferred compounds according to the present invention include:

-   -   Compounds of formula I, IA, IB, IC, ID, IF, IG, Ia, IAa, IBa,        ICa, IDa, IFa, IGa, Ib, IAb, IBb, ICb, IDb, IFb, and IGb        wherein:    -   R₄ is selected from —CH₂OH and —CH₂OC(═O)R^(c);    -   Being particularly preferred compounds of formula Ia, IAa, IBa,        ICa, IDa, IFa, and IGa and/or compounds where R₄ is —CH₂OH.    -   Compounds of formula I, IA, IB, IC, ID, IE IF, IG, Ia, IAa, IBa,        ICa, IDa, IEa, IFa, IGa, Ib, IAb, IBb, ICb, IDb, IEb, IFb, and        IGb wherein    -   R₄ is selected from —CH₂NH₂ and —CH₂NHProt^(NH); and    -   Prot^(NH) is a protecting group for amino.    -   Being particularly preferred compounds of formula Ib, IAb, IBb,        ICb, IDb, IEb, IFb, and IGb and/or compounds where R₄ is        —CH₂NH₂.    -   Compounds of formula Ic, IAc, IBc, IDc, IFc, IGc wherein    -   R₂ is a —C(═O)R^(a) group for compounds of formula Ic, IAc, IBc,        IDc, or IFc; and R₂ is acetyl for compounds of formula IGc;    -   R₃ is hydrogen or a —OR^(b) group for compounds of formula Ic,        IDc, IFc, IGc; R₃ is hydrogen for compounds of formula IAc; or        R₃ is a —OR^(b) group for compounds of formula IBc;    -   R^(a) is selected from hydrogen, and substituted or        unsubstituted C₁-C₆ alkyl; and    -   R^(b) is substituted or unsubstituted C₁-C₆ alkyl.

Particularly preferred compounds according to the present inventioninclude:

-   -   Compounds of formula I, IA, IB, IC, IF, IG, Ia, IAa, IBa, ICa,        IFa, IGa, Ib, IAb, IBb, ICb, IFb, and IGb wherein    -   X is —NH—;    -   R₄ is selected from —CH₂OH, and —CH₂OC(═O)R^(c);    -   and    -   R^(c) is selected from substituted or unsubstituted C₁-C₁₂        alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl, and        substituted or unsubstituted C₂-C₁₂ alkynyl.    -   Being more preferred compounds of formula Ia, IAa, IBa, ICa,        IFa, IGa and/or compounds where R₄ is —CH₂OH.    -   Compounds of formula I, IA, IB, ID, IF, IG, Ia, IAa, IBa, IDa,        IFa, IGa, Ib, IAb, IBb, IDb, IFb, and IGb wherein    -   X is —O—;    -   R₄ is selected from —CH₂OH and —CH₂OC(═O)R^(c);    -   and is selected from substituted or unsubstituted C₁-C₁₂ alkyl,        substituted or unsubstituted C₂-C₁₂ alkenyl, and substituted or        unsubstituted C₂-C₁₂ alkynyl.    -   Being more preferred compounds of formula Ia, IAa, IBa, IDa,        IFa, IGa and/or compounds where R₄ is —CH₂OH.    -   Compounds of formula I, IA, IB, IC, IE, IF, IG, Ia, IAa, IBa,        ICa, IEa, IFa, IGa, Ib, IAb, IBb, ICb, IEb, IFb, and IGb wherein    -   X is —NH—;    -   R₄ is selected from —CH₂NH₂ and —CH₂NHProt^(NH);    -   and    -   Prot^(NH) is a protecting group for amino.    -   Being more preferred compounds of formula Ib, IAb, IBb, ICb,        IEb, IFb, IGb and/or compounds where R₄ is —CH₂NH₂.    -   Compounds of formula I, IA, IB, ID, IE, IF, IG, Ia, IAa, IBa,        IDa, IEa, IFa, IGa, Ib, IAb, IBb, IDb, IEb, IFb, and IGb wherein    -   X is —O—;    -   R₄ is selected from —CH₂NH₂ and —CH₂NHProt^(NH);    -   and    -   Prot^(NH) is a protecting group for amino.    -   Being more preferred compounds of formula Ib, IAb, IBb, IDb,        IEb, IFb, IGb and/or compounds where R₄ is —CH₂NH₂.    -   Compounds of formula I, IA, IB, IC, ID, IF, IG, Ia, IAa, IBa,        ICa, IDa, IFa, IGa, Ib, IAb, IBb, ICb, IDb, IFb, IGb wherein    -   R₂ is a —C(═O)R^(a) group for compounds of formula I, IA, IB,        IC, ID, IF, Ia, IAa, IBa, ICa, IDa, IFa, Ib, IAb, IBb, ICb, IDb,        or IFb; and R₂ is acetyl for compounds of formula IG, IGa or        IGb;    -   R₃ is hydrogen or a —OR^(b) group for compounds of formula I,        IC, ID, IF, IG, Ia, ICa, IDa, IFa, IGa, Ib, ICb, IDb, IFb, or        IGb; R₃ is hydrogen for compounds of formula    -   IA, IAa or IAb; or R₃ is a —OR^(b) group for compounds of        formula IB, IBa or IBb;    -   R₄ is selected from —CH₂OH, and —CH₂OC(═O)R^(c);    -   R^(a) is selected from hydrogen, and substituted or        unsubstituted C₁-C₆ alkyl;    -   R^(b) is substituted or unsubstituted C₁-C₆ alkyl; and    -   R^(c) is substituted or unsubstituted C₁-C₆ alkyl.    -   Being more preferred compounds of formula Ia, IAa, IBa, ICa,        IDa, IFa, IGa and/or compounds where R₄ is —CH₂OH.    -   Compounds of formula I, IA, IB, IC, ID, IE, IF, IG, Ia, IAa,        IBa, ICa, IDa, IEa, IFa, IGa, Ib, IAb, IBb, ICb, IDb, IEb, IFb,        and IGb wherein    -   R₂ is a —C(═O)R^(a) group for compounds of formula I, IA, IB,        IC, ID, IF, Ia, IAa, IBa,    -   ICa, IDa, IFa, Ib, IAb, IBb, ICb, IDb, or IFb; and R₂ is acetyl        for compounds of formula IG, IGa or IGb;    -   R₃ is hydrogen or a —OR^(b) group for compounds of formula I,        IC, ID, IE, IF, IG, Ia, ICa, IDa, IEa, IFa, IGa, Ib, ICb, IDb,        IEb, IFb, or IGb; R₃ is hydrogen for compounds of formula IA,        IAa or IAb; or R₃ is a —OR^(b) group for compounds of formula        IB, IBa or IBb;    -   R₄ is selected from —CH₂NH₂ and —CH₂NHProt^(NH);    -   R^(a) is selected from hydrogen, and substituted or        unsubstituted C₁-C₆ alkyl;    -   R^(b) is substituted or unsubstituted C₁-C₆ alkyl; and    -   Prot^(NH) is a protecting group for amino.    -   Being more preferred compounds of formula Ib, IAb, IBb, ICb,        IDb, IEb, IFb, IGb and/or compounds where R₄ is —CH₂NH₂    -   Compounds of formula Ic, IAc, IBc, IDc, IFc, IGc wherein    -   R₂ is a —C(═O)R^(a) group for compounds of formula Ic, IAc, IBc,        IDc, or IFc; and R₂ is acetyl for compounds of formula IGc;    -   R₃ is hydrogen or a —OR^(b) group for compounds of formula Ic,        IDc, IFc, IGc; R₃ is hydrogen for compounds of formula IAc; or        R₃ is a —OR^(b) group for compounds of formula IBc;    -   R^(a) is substituted or unsubstituted C₁-C₆ alkyl; and    -   R^(b) is substituted or unsubstituted C₁-C₆ alkyl.    -   More preferred compounds according to the present invention        include    -   Compounds of formula I, IA, IB, IC, IF, IG, Ia, IAa, IBa, ICa,        IFa, IGa, Ib, IAb, IBb, ICb, IFb, and IGb wherein    -   X is —NH—;    -   R₂ is a —C(═O)R^(a) group for compounds of formula I, IA, IB,        IC, IF, Ia, IAa, IBa, ICa, IFa, Ib, IAb, IBb, ICb, or IFb; and        R₂ is acetyl for compounds of formula IG, IGa or    -   IGb;    -   R₃ is hydrogen or a —OR^(b) group for compounds of formula I,        IC, IF, IG, Ia, ICa, IFa, IGa, Ib, ICb, IFb, or IGb; R₃ is        hydrogen for compounds of formula IA, IAa or IAb; or R₃ is a        —OR^(b) group for compounds of formula IB, IBa or IBb;    -   R₄ is —CH₂OH;    -   R^(a) is selected from hydrogen and substituted or unsubstituted        C₁-C₆ alkyl; and    -   R^(b) is substituted or unsubstituted C₁-C₆ alkyl.    -   Being particularly more preferred compounds of formula Ia, IAa,        or IBa, ICa, IFa, IGa.    -   Compounds of formula I, IA, IB, ID, IF, IG, Ia, IAa, IBa, IDa,        IFa, IGa, Ib, IAb, IBb, IDb, IFb, and IGb wherein    -   X is —O—;    -   R₂ is a —C(═O)R^(a) group for compounds of formula I, IA, IB,        ID, IF, Ia, IAa, IBa, IDa, IFa, Ib, IAb, IBb, IDb, or IFb; and        R₂ is acetyl for compounds of formula IG,    -   IGa or IGb;    -   R₃ is hydrogen or a —OR^(b) group for compounds of formula I,        ID, IF, IG, Ia, IDa, IFa, IGa, Ib, IDb, IFb, or IGb; R₃ is        hydrogen for compounds of formula IA, IAa or IAb; or R₃ is a        —OR^(b) group for compounds of formula IB, IBa or IBb;    -   R₄ is —CH₂OH;    -   R^(a) is selected from hydrogen and substituted or unsubstituted        C₁-C₆ alkyl; and    -   R^(b) is substituted or unsubstituted C₁-C₆ alkyl.    -   Being particularly more preferred compounds of formula Ia, IAa,        IBa, IDa, IFa, or IGa.    -   Compounds of formula I, IA, IB, IC, IE, IF, IG, Ia, IAa, IBa,        ICa, IEa, IFa, IGa,    -   Ib, IAb, IBb, ICb, IEb, IFb, and IGb wherein    -   X is —NH—;    -   R₂ is a —C(═O)R^(a) group for compounds of formula I, IA, IB,        IC, IE, IF, Ia, IAa, IBa, ICa, IEa, IFa, Ib, IAb, IBb, ICb, IEb        or IFb; and R₂ is acetyl for compounds of formula IG, IGa or        IGb;    -   R₃ is hydrogen or a —OR^(b) group for compounds of formula I,        IC, IE, IF, IG, Ia, ICa, IEa, IFa, IGa, Ib, ICb, IEb, IFb, or        IGb; R₃ is hydrogen for compounds of formula IA, IAa or IAb; or        R₃ is a —OR^(b) group for compounds of formula IB, IBa or IBb;    -   R₄ is selected from —CH₂NH₂ and —CH₂NHProt^(NH);    -   R^(a) is selected from hydrogen and substituted or unsubstituted        C₁-C₆ alkyl;    -   R^(b) is substituted or unsubstituted C₁-C₆ alkyl; and    -   Prot^(NH) is a protecting group for amino.    -   Being particularly more preferred compounds of formula Ib, IAb,        IBb, ICb, IEb, IFb, IGb and/or compounds where R₄ is —CH₂NH₂    -   Compounds of formula I, IA, IB, ID, IE, IF, IG, Ia, IAa, IBa,        IDa, IEa, IFa, IGa,    -   Ib, IAb, IBb, IDb, IEb, IFb, and IGb wherein    -   X is —O—;    -   R₂ is a —C(═O)R^(a) group for compounds of formula I, IA, IB,        ID, IE, IF, Ia, IAa, IBa, IDa, IEa, IFa, Ib, IAb, IBb, IDb, IEb        or IFb; and R₂ is acetyl for compounds of formula IG, IGa or        IGb;    -   R₃ is hydrogen or a —OR^(b) group for compounds of formula I,        ID, IE, IF, IG, Ia, IDa,    -   IEa, IFa, IGa, Ib, IDb, IEb, IFb, or IGb; R₃ is hydrogen for        compounds of formula IA, IAa or IAb; or R₃ is a —OR^(b) group        for compounds of formula IB, IBa or IBb;    -   R₄ is selected from —CH₂NH₂ and —CH₂NHProt^(NH);    -   R^(a) is selected from hydrogen and substituted or unsubstituted        C₁-C₆ alkyl;    -   R^(b) is substituted or unsubstituted C₁-C₆ alkyl; and    -   Prot^(NH) is a protecting group for amino.    -   Being particularly more preferred compounds of formula Ib, IAb,        IBb, IDb, IEb, IFb, IGb and/or compounds where R₄ is CH₂NH₂.    -   Compounds of formula I, IA, IB, IC, ID, IF, IG, Ia, IAa, IBa,        ICa, IDa, IFa, IGa, Ib, IAb, IBb, ICb, IDb, IFb, and IGb wherein    -   R₂ is a —C(═O)R^(a) group for compounds of formula I, IA, IB,        IC, ID, IF, Ia, IAa, IBa, ICa, IDa, IFa, Ib, IAb, IBb, ICb, IDb        or IFb; and R₂ is acetyl for compounds of formula IG, IGa or        IGb;    -   R₃ is hydrogen or a —OR^(b) group for compounds of formula I,        IC, ID, IF, IG, Ia, ICa, IDa, IFa, IGa, Ib, ICb, IDb, IFb, or        IGb; R₃ is hydrogen for compounds of formula    -   IA, IAa or IAb; or R₃ is a —OR^(b) group for compounds of        formula IB, IBa or IBb;    -   R₄ is —CH₂OH;    -   R^(a) is substituted or unsubstituted C₁-C₆ alkyl; and    -   R^(b) is substituted or unsubstituted C₁-C₆ alkyl.    -   Being particularly more preferred compounds of formula Ia, IAa,        IBa, ICa, IDa,    -   IFa, or IGa.    -   Compounds of formula I, IA, IB, IC, ID, IE, IF, IG, Ia, IAa,        IBa, ICa, IDa, IEa, IFa, IGa, Ib, IAb, IBb, ICb, IDb, IEb, IFb,        and IGb wherein    -   R₂ is a —C(═O)R^(a) group for compounds of formula I, IA, IB,        IC, ID, IE, IF, Ia, IAa, IBa, ICa, IDa, IEa, IFa, Ib, IAb, IBb,        ICb, IDb, IEb or IFb; and R₂ is acetyl for compounds of formula        IG, IGa or IGb;    -   R₃ is hydrogen or a —OR^(b) group for compounds of formula I,        IC, ID, IE, IF, IG, Ia, ICa, IDa, IEa, IFa, IGa, Ib, ICb, IDb,        IEb, IFb, or IGb; R₃ is hydrogen for compounds of formula IA,        IAa or IAb; or R₃ is a —OR^(b) group for compounds of formula        IB, IBa or IBb;    -   R₄ is selected from —CH₂NH₂ and —CH₂NHProt^(NH);    -   R^(a) is substituted or unsubstituted C₁-C₆ alkyl;    -   R^(b) is substituted or unsubstituted C₁-C₆ alkyl; and    -   Prot^(NH) is a protecting group for amino.    -   Being particularly more preferred compounds of formula Ib, IAb,        IBb, ICb, IDb, IEb, IFb, IGb and/or compounds where R₄ is        —CH₂NH₂.    -   Compounds of formula I, IA, IB, IC, IF, IG, Ia, IAa, IBa, ICa,        IFa, IGa, Ib, IAb, IBb, ICb, IFb, and IGb wherein    -   X is —NH—;    -   R₂ is a —C(═O)R^(a) group for compounds of formula I, IA, IB,        IC, IF, Ia, IAa, IBa, ICa, IFa, Ib, IAb, IBb, ICb, or IFb; and        R₂ is acetyl for compounds of formula IG, IGa or    -   IGb;    -   R₃ is hydrogen or a —OR^(b) group for compounds of formula I,        IC, IF, IG, Ia, ICa, IFa, IGa, Ib, ICb, IFb, or IGb; R₃ is        hydrogen for compounds of formula IA, IAa or IAb; or R₃ is a        —OR^(b) group for compounds of formula IB, IBa or IBb;    -   R₄ is —CH₂OC(═O)R^(c);    -   R^(a) is selected from hydrogen and substituted or unsubstituted        C₁-C₆ alkyl;    -   R^(b) is substituted or unsubstituted C₁-C₆ alkyl; and    -   is a substituted or unsubstituted C₁-C₆ alkyl.    -   Being more preferred compounds of formula Ia, IAa, IBa, ICa,        IFa, or IGa.    -   Compounds of formula Ic, IAc, IBc, IDc, IFc, and IGc wherein    -   R₂ is a —C(═O)R^(a) group for compounds of formula Ic, IAc, IBc,        IDc, or IFc; and R₂ is acetyl for compounds of formula IGc;    -   R₃ is hydrogen or methoxy for compounds of formula Ic, IDc, IFc,        or IGc; R₃ is hydrogen for compounds of formula IAc; or R₃ is        methoxy for compounds of formula IBc; and    -   R^(a) is substituted or unsubstituted C₁-C₆ alkyl.    -   Particularly more preferred compounds according to the present        invention include:    -   Compounds of formula I, IA, IB, IC, IF, IG, Ia, IAa, IBa, ICa,        IFa, IGa, Ib, IAb, IBb, ICb, IFb, and IGb wherein    -   X is —NH—;    -   R₂ is a —C(═O)R^(a) group for compounds of formula I, IA, IB,        IC, IF, Ia, IAa, IBa, ICa,    -   IFa, Ib, IAb, IBb, ICb, or IFb; and R₂ is acetyl for compounds        of formula IG, IGa or IGb;    -   R₃ is hydrogen or methoxy for compounds of formula I, IC, IF,        IG, Ia, ICa, IFa, IGa, Ib, ICb, IFb, or IGb; R₃ is hydrogen for        compounds of formula IA, IAa or IAb; and    -   R₃ is methoxy for compounds of formula IB, IBa or IBb;    -   R₄ is —CH₂OH; and    -   R^(a) is substituted or unsubstituted C₁-C₆ alkyl.    -   Being even more preferred compounds of formula Ia, IAa, IBa,        ICa, IFa, IGa.    -   Compounds of formula I, IA, IB, ID, IF, IG, Ia, IAa, IBa, IDa,        IFa, IGa, Ib, IAb, IBb, IDb, IFb, and IGb wherein    -   X is —O—;    -   R₂ is a —C(═O)R^(a) group for compounds of formula I, IA, IB,        ID, IF, Ia, IAa, IBa, IDa, IFa, Ib, IAb, IBb, IDb, or IFb; and        R₂ is acetyl for compounds of formula IG, IGa or IGb;    -   R₃ is hydrogen or methoxy for compounds of formula I, ID, IF,        IG, Ia, IDa, IFa, IGa,    -   Ib, IDb, IFb, or IGb; R₃ is hydrogen for compounds of formula        IA, IAa or IAb; or R₃ is methoxy for compounds of formula IB,        IBa or IBb;    -   R₄ is —CH₂OH; and    -   R^(a) is substituted or unsubstituted C₁-C₆ alkyl.    -   Being even more preferred compounds of formula Ia, IAa, IBa,        IDa, IEa, IFa, IGa.    -   Compounds of formula I, IA, IB, IC, IE, IF, IG, Ia, IAa, IBa,        ICa, IEa, IFa, IGa, Ib, IAb, IBb, ICb, IEb, IFb, and IGb wherein    -   X is —NH—;    -   R₂ is a —C(═O)R^(a) group for compounds of formula I, IA, IB,        IC, IE, IF, Ia, IAa, IBa, ICa, IEa, IFa, Ib, IAb, IBb, ICb, IEb        or IFb; and R₂ is acetyl for compounds of formula IG, IGa or        IGb;    -   R₃ is hydrogen or methoxy for compounds of formula I, IC, IE,        IF, IG, Ia, ICa, IEa, IFa, IGa, Ib, ICb, IEb, IFb, or IGb; R₃ is        hydrogen for compounds of formula IA, IAa or IAb; or R₃ is        methoxy for compounds of formula IB, IBa or IBb;    -   R₄ is selected from —CH₂NH₂ and —CH₂NHProt^(NH);    -   R^(a) is substituted or unsubstituted C₁-C₆ alkyl; and    -   Prot^(NH) is a protecting group for amino.    -   Being even more preferred compounds of formula Ib, IAb, IBb,        ICb, IEb, IFb, IGb and/or compounds where R₄ is —CH₂NH₂.    -   Compounds of formula I, IA, IB, ID, IE, IF, IG, Ia, IAa, IBa,        IDa, IEa, IFa, IGa, Ib, IAb, IBb, IDb, IEb, IFb, and IGb wherein    -   X is —O—;    -   R₂ is a —C(═O)R^(a) group for compounds of formula I, IA, IB,        ID, IE, IF, Ia, IAa, IBa,    -   IDa, IEa, IFa, Ib, IAb, IBb, IDb, IEb or IFb; and R₂ is acetyl        for compounds of formula IG, IGa or IGb;    -   R₃ is hydrogen or methoxy for compounds of formula I, ID, IE,        IF, IG, Ia, IDa, IEa, IFa, IGa, Ib, IDb, IEb, IFb, or IGb; R₃ is        hydrogen for compounds of formula IA, IAa or IAb; or R₃ is        methoxy for compounds of formula IB, IBa or IBb;    -   R₄ is selected from —CH₂NH₂ and —CH₂NHProt^(NH);    -   R^(a) is substituted or unsubstituted C₁-C₆ alkyl; and    -   Prot^(NH) is a protecting group for amino.    -   Being even more preferred compounds of formula Ib, IAb, IBb,        IDb, IEb, IFb, IGb and/or compounds where R₄ is —CH₂NH₂.    -   Compounds of formula I, IA, IB, IC, ID, IF, IG, Ia, IAa, IBa,        ICa, IDa, IFa, IGa, Ib, IAb, IBb, ICb, IDb, IFb, and IGb wherein    -   R₂ is a —C(═O)R^(a) group for compounds of formula I, IA, IB,        IC, ID, IF, Ia, IAa, IBa, ICa, IDa, IFa, Ib, IAb, IBb, ICb, IDb,        or IFb; and R₂ is acetyl for compounds of formula IG, IGa or        IGb;    -   R₃ is hydrogen or methoxy for compounds of formula I, IC, ID,        IF, IG, Ia, ICa, IDa, IFa, IGa, Ib, ICb, IDb, IFb, and IGb; R₃        is hydrogen for compounds of formula IA, IAa or IAb; or R₃ is        methoxy for compounds of formula IB, IBa or IBb;    -   R₄ is —CH₂OH; and    -   R^(a) is selected from methyl, ethyl, n-propyl, isopropyl and        butyl, including n-butyl, sec-butyl, isobutyl and tert-butyl.    -   Being even more preferred compounds of formula Ia, IAa, IBa,        ICa, IDa, IEa, IFa, or IGa.    -   Compounds of formula I, IA, IB, IC, ID, IE, IF, IG, Ia, IAa,        IBa, ICa, IDa, IEa, IFa, IGa, Ib, IAb, IBb, ICb, IDb, IEb, IFb,        and IGb wherein    -   R₂ is a —C(═O)R^(a) group for compounds of formula I, IA, IB,        IC, ID, IE, IF, Ia, IAa, IBa, ICa, IDa, IEa, IFa, Ib, IAb, IBb,        ICb, IDb, IEb or IFb; and R₂ is acetyl for compounds of formula        IG, IGa or IGb;    -   R₃ is hydrogen or a methoxy for compounds of formula I, IC, ID,        IE, IF, IG, Ia, ICa, IDa, IEa, IFa, IGa, Ib, ICb, IDb, IEb, IFb,        and IGb; R₃ is hydrogen for compounds of formula IA, IAa or IAb;        or R₃ is methoxy for compounds of formula IB, IBa or IBb;    -   R₄ is selected from —CH₂NH₂ and —CH₂NHProt^(NH);    -   R^(a) is selected from methyl, ethyl, n-propyl, isopropyl and        butyl, including n-butyl, sec-butyl, isobutyl and tert-butyl;        and    -   Prot^(NH) is a protecting group for amino.    -   Being even more preferred compounds of formula Ib, IAb, IBb,        ICb, IDb, IEb, IFb, IGb and/or compounds where R₄ is —CH₂NH₂.    -   Compounds of formula Ic or IAc, IDc, IFc, and IGc wherein    -   R₂ is a —C(═O)R^(a) group for compounds of formula Ic, IAc, IDc,        or IFc; and R₂ is acetyl for compounds of formula IGc;    -   R₃ is hydrogen; and    -   R^(a) is selected from methyl, ethyl, n-propyl, isopropyl and        butyl, including n-butyl, sec-butyl, isobutyl and tert-butyl.    -   Compounds of formula Ic, IBc, IDc, IFc, and IGc wherein    -   R₂ is a —C(═O)R^(a) group for compounds of formula Ic, IBc, IDc,        or IFc; and R₂ is acetyl for compounds of formula IGc;    -   R₃ is methoxy; and    -   R^(a) is selected from methyl, ethyl, n-propyl, isopropyl and        butyl, including n-butyl, sec-butyl, isobutyl and tert-butyl.

Even more preferred compounds according to the present inventioninclude:

-   -   Compounds of formula I, IA, IC, IF, IG, Ia, IAa, ICa, IFa, IGa,        Ib, IAb, ICb, IFb, and IGb wherein    -   X is —NH—;    -   R₂ is acetyl;    -   R₃ is hydrogen; and    -   R₄ is —CH₂OH.    -   Being most preferred compounds of formula Ia, IAa, ICa, IFa, or        IGa.    -   Compounds of formula I, IA, ID, IF, IG, Ia, IAa, IDa, IFa, IGa,        Ib, IAb, IDb, IFb, and IGb wherein    -   X is —O—;    -   R₂ is acetyl;    -   R₃ is hydrogen; and    -   R₄ is —CH₂OH.    -   Being most preferred compounds of formula Ia, IAa, IDa, IFa, or        IGa    -   Compounds of formula I, IA, IC, IE, IF, IG, Ia, IAa, ICa, IEa,        IFa, IGa, Ib, IAb, ICb, IEb, IFb, and IGb wherein    -   X is —NH—;    -   R₂ is acetyl;    -   R₃ is hydrogen; and    -   R₄ is —CH₂NH₂.    -   Being most preferred compounds of formula Ib, IAb, ICb, IEb,        IFb, or IGb.    -   Compounds of formula I, IA, ID, IE, IF, IG, Ia, IAa, IDa, IEa,        IFa, IGa, Ib, IAb, IDb, IEb, IFb, and IGb wherein    -   X is —O—;    -   R₂ is acetyl;    -   R₃ is hydrogen; and    -   R₄ is —CH₂NH₂.    -   Being most preferred compounds of formula Ib, IAb, IDb, IEb,        IFb, or IGb.    -   Compounds of formula I, IA, IC, ID, IF, IG, Ia, IAa, ICa, IDa,        IFa, IGa, Ib, IAb, ICb, IDb, IFb, and IGb wherein    -   R₂ is acetyl;    -   R₃ is hydrogen; and    -   R₄ is —CH₂OH.    -   Being most preferred compounds of formula Ia, IAa, ICa, IDa, IFa        or IGa.    -   Compounds of formula I, IA, IC, ID, IF, IG, Ia, IAa, ICa, IDa,        IFa, IGa, Ib, IAb, ICb, IDb, IFb, and IGb wherein    -   R₁ is —OH;    -   R₂ is acetyl;    -   R₃ is hydrogen; and    -   R₄ is —CH₂OH.    -   Being most preferred compounds of formula Ia, IAa, ICa, IDa, IFa        or IGa.    -   Compounds of formula I, IA, IC, ID, IE, IF, IG, Ia, IAa, ICa,        IDa, IEa, IFa, IGa, Ib, IAb, ICb, IDb, IEb, IFb, and IGb wherein    -   R₂ is acetyl;    -   R₃ is hydrogen; and    -   R₄ is —CH₂NH₂.    -   Being most preferred compounds of formula Ib, IAb, ICb, IDb,        IEb, IFb, or IGb.    -   Compounds of formula Ic or IAc, IDc, IFc, IGc wherein    -   R₂ is acetyl; and    -   R₃ is hydrogen.    -   Compounds of formula Ic or IBc, IDc, IFc, IGc wherein    -   R₂ is acetyl; and    -   R₃ is methoxy.    -   A compound according to the present invention of formula:

-   -   or a pharmaceutically acceptable salt or ester thereof.    -   Being particularly preferred a compound of formula:

-   -   or a pharmaceutically acceptable salt or ester thereof    -   A compound according to the present invention of formula:

-   -   or a pharmaceutically acceptable salt or ester thereof.    -   Being particularly preferred a compound of formula:

-   -   or a pharmaceutically acceptable salt or ester thereof.    -   Being more preferred a compound of formula:

-   -   or a pharmaceutically acceptable salt or ester thereof.        Being even more preferred compounds according to the present        invention are compounds of formula:

or a pharmaceutically acceptable salt or ester thereof.The most preferred compounds according to the present invention arecompounds of formula:

or a pharmaceutically acceptable salt or ester thereof.

In additional preferred embodiments, the preferences described above forthe different substituents are combined. The present invention is alsodirected to such combinations of preferred substitutions (where allowedby possible substituent groups) in compounds of formula I, IA, IB, IC,ID, IE, IF, IG, Ia, IAa, IBa, ICa, IDa, IEa, IFa, IGa, Ib, IAb, IBb,ICb, IDb, IEb, IFb, IGb, Ic, IAc, IBc, IDc, IFc or IGc according to thepresent invention.

An important feature of the above-described compounds is theirbioactivity and in particular their cytotoxic activity. In this regard,we have surprisingly found that the compounds of the present inventionshow an enhanced antitumor activity, as it is shown in Examples 27 and29 to 40.

Compositions Comprising a Compound of Formula I, IA, IB, IC, ID, IE, IF,IG, Ia, IAa, IBa, ICa, IDa, IEa, IFa, IGa, Ib, IAb, IBb, ICb, IDb, IEb,IFb, IGb, Ic, IAc, IBc, IDc, IFc or IGc of the Invention and UsesThereof

In a further embodiment of the present invention, there is provided apharmaceutical composition comprising a compound according to thepresent invention and a pharmaceutically acceptable carrier. Examples ofthe administration form include without limitation oral, topical,parenteral, sublingual, rectal, vaginal, ocular and intranasal.Parenteral administration includes subcutaneous injections, intravenous,intramuscular, intrasternal injection or infusion techniques. Preferablythe compositions are administered parenterally. Pharmaceuticalcompositions of the invention can be formulated so as to allow acompound according to the present invention to be bioavailable uponadministration of the composition to an animal, preferably human.Compositions can take the form of one or more dosage units, where forexample, a tablet can be a single dosage unit, and a container of acompound according to the present invention may contain the compound inliquid or in aerosol form and may hold a single or a plurality of dosageunits.

The pharmaceutically acceptable carrier or vehicle can be particulate,so that the compositions are, for example, in tablet or powder form. Thecarrier(s) can be liquid, with the compositions being, for example, anoral syrup or injectable liquid. In addition, the carrier(s) can begaseous, or liquid so as to provide an aerosol composition useful in,for example inhalatory administration. Powders may also be used forinhalation dosage forms. The term “carrier” refers to a diluent,adjuvant or excipient, with which the compound according to the presentinvention is administered. Such pharmaceutical carriers can be liquids,such as water and oils including those of petroleum, animal, vegetableor synthetic origin, such as peanut oil, soybean oil, mineral oil,sesame oil and the like. The carriers can be saline, gum acacia,gelatin, starch paste, talc, keratin, colloidal silica, urea,disaccharides, and the like. In addition, auxiliary, stabilizing,thickening, lubricating and coloring agents can be used. In oneembodiment, when administered to an animal, the compounds andcompositions according to the present invention, and pharmaceuticallyacceptable carriers are sterile. Water is a preferred carrier when thecompounds according to the present invention are administeredintravenously. Saline solutions and aqueous dextrose and glycerolsolutions can also be employed as liquid carriers, particularly forinjectable solutions. Suitable pharmaceutical carriers also includeexcipients such as starch, glucose, lactose, sucrose, gelatin, malt,rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate,talc, sodium chloride, dried skim milk, glycerol, propylene glycol,water, ethanol and the like. The present compositions, if desired, canalso contain minor amounts of wetting or emulsifying agents, or pHbuffering agents.

When intended for oral administration, the composition is preferably insolid or liquid form, where semi-solid, semi-liquid, suspension and gelforms are included within the forms considered herein as either solid orliquid.

As a solid composition for oral administration, the composition can beformulated into a powder, granule, compressed tablet, pill, capsule,chewing gum, wafer or the like form. Such a solid composition typicallycontains one or more inert diluents. In addition, one or more for thefollowing can be present: binders such as carboxymethylcellulose, ethylcellulose, microcrystalline cellulose, or gelatin; excipients such asstarch, lactose or dextrins, disintegrating agents such as alginic acid,sodium alginate, corn starch and the like; lubricants such as magnesiumstearate; glidants such as colloidal silicon dioxide; sweetening agentsuch as sucrose or saccharin; a flavoring agent such as peppermint,methyl salicylate or orange flavoring; and a coloring agent.

When the composition is in the form of a capsule (e.g. a gelatincapsule), it can contain, in addition to materials of the above type, aliquid carrier such as polyethylene glycol, cyclodextrins or a fattyoil.

The composition can be in the form of a liquid, e.g. an elixir, syrup,solution, emulsion or suspension. The liquid can be useful for oraladministration or for delivery by injection. When intended for oraladministration, a composition can comprise one or more of a sweeteningagent, preservatives, dye/colorant and flavor enhancer. In a compositionfor administration by injection, one or more of a surfactant,preservative, wetting agent, dispersing agent, suspending agent, buffer,stabilizer and isotonic agent can also be included.

The preferred route of administration is parenteral administrationincluding, but not limited to, intradermal, intramuscular,intraperitoneal, intravenous, subcutaneous, intranasal, epidural,intracerebral, intraventricular, intrathecal, intravaginal ortransdermal. The preferred mode of administration is left to thediscretion of the practitioner, and will depend in part upon the site ofthe medical condition (such as the site of cancer). In a more preferredembodiment, the compounds according to the present invention areadministered intravenously. Infusion times of up to 24 hours arepreferred to be used, more preferably 1 to 12 hours, with 1 to 6 hoursbeing most preferred. Short infusion times which allow treatment to becarried out without an overnight stay in a hospital are especiallydesirable. However, infusion may be 12 to 24 hours or even longer ifrequired. Infusion may be carried out at suitable intervals of, forexample, 1 to 4 weeks.

The liquid compositions of the invention, whether they are solutions,suspensions or other like form, can also include one or more of thefollowing: sterile diluents such as water for injection, salinesolution, preferably physiological saline, Ringer's solution, isotonicsodium chloride, fixed oils such as synthetic mono or diglycerides,polyethylene glycols, glycerin, or other solvents; antibacterial agentssuch as benzyl alcohol or methyl paraben; and agents for the adjustmentof tonicity such as sodium chloride or dextrose. A parenteralcomposition can be enclosed in an ampoule, a disposable syringe or amultiple-dose vial made of glass, plastic or other material.Physiological saline is a preferred adjuvant.

The amount of the compound according to the present invention that iseffective in the treatment of a particular disorder or condition willdepend on the nature of the disorder or condition, and can be determinedby standard clinical techniques. In addition, in vitro or in vivo assayscan optionally be employed to help identify optimal dosage ranges. Theprecise dose to be employed in the compositions will also depend on theroute of administration, and the seriousness of the disease or disorder,and should be decided according to the judgement of the practitioner andeach patient's circumstances.

The compositions comprise an effective amount of a compound of thepresent invention such that a suitable dosage will be obtained. Thecorrect dosage of the compounds will vary according to the particularformulation, the mode of application, and its particular site, host andthe disease being treated, e.g. cancer and, if so, what type of tumor.Other factors like age, body weight, sex, diet, time of administration,rate of excretion, condition of the host, drug combinations, reactionsensitivities and severity of the disease should be taken into account.Administration can be carried out continuously or periodically withinthe maximum tolerated dose.

Typically, the amount is at least about 0.01% of a compound of thepresent invention, and may comprise at least 80%, by weight of thecomposition. When intended for oral administration, this amount can bevaried to range from about 0.1% to about 80% by weight of thecomposition. Preferred oral compositions can comprise from about 4% toabout 50% of the compound of the present invention by weight of thecomposition.

Preferred compositions of the present invention are prepared so that aparenteral dosage unit contains from about 0.01% to about 10% by weightof the compound of the present invention. More preferred parenteraldosage unit contains about 0.5% to about 5% by weight of the compound ofthe present invention.

For intravenous administration, the composition is suitable for dosesfrom about 0.1 mg/kg to about 250 mg/kg of the animal's body weight,preferably from about 0.1 mg/kg and about 20 mg/kg of the animal's bodyweight, and more preferably from about 1 mg/kg to about 10 mg/kg of theanimal's body weight.

The compound of the present invention, can be administered by anyconvenient route, for example by infusion or bolus injection, byabsorption through epithelial or mucocutaneous linings.

In specific embodiments, it can be desirable to administer one or morecompounds of the present invention, or compositions locally to the areain need of treatment. In one embodiment, administration can be by directinjection at the site (or former site) of a cancer, tumor or neoplasticor pre-neoplastic tissue.

Pulmonary administration can also be employed, e.g. by use of an inhaleror nebulizer, and formulation with an aerosolizing agent, or viaperfusion in a fluorocarbon or synthetic pulmonary surfactant. Incertain embodiments, the compound of the present invention can beformulated as a suppository, with traditional binders and carriers suchas triglycerides.

The present compositions can take the form of solutions, suspensions,emulsions, tablets, pills, pellets, capsules, capsules containingliquids, powders, sustained-release formulations, suppositories,emulsions, aerosols, sprays, suspensions, or any other form suitable foruse. Other examples of suitable pharmaceutical carriers are described in“Remington's Pharmaceutical Sciences” by E. W. Martin.

The pharmaceutical compositions can be prepared using methodology wellknown in the pharmaceutical art. For example, a composition intended tobe administered by injection can be prepared by combining a compound ofthe present invention with water, or other physiologically suitablediluent, such as phosphate buffered saline, so as to form a solution. Asurfactant can be added to facilitate the formation of a homogeneoussolution or suspension.

Preferred compositions according to the present invention include:

-   -   Pharmaceutical compositions comprising a compound of the present        invention and a disaccharide. Particularly preferred        disaccharides are selected from lactose, trehalose, sucrose,        maltose, isomaltose, cellobiose, isosaccharose, isotrehalose,        turanose, melibiose, gentiobiose, and mixtures thereof    -   Lyophilised pharmaceutical compositions comprising a compound of        the present invention and a disaccharide. Particularly preferred        disaccharides are selected from lactose, trehalose, sucrose,        maltose, isomaltose, cellobiose, isosaccharose, isotrehalose,        turanose, melibiose, gentiobiose, and mixtures thereof.

The ratio of the active substance to the disaccharide in embodiments ofthe present invention is determined according to the solubility of thedisaccharide and, when the formulation is freeze dried, also accordingto the freeze-dryability of the disaccharide. It is envisaged that thisactive substance:disaccharide ratio (w/w) can be about 1:10 in someembodiments, about 1:20 in other embodiments, about 1:50 in still otherembodiments. It is envisaged that other embodiments have such ratios inthe range from about 1:5 to about 1:500, and still further embodimentshave such ratios in the range from about 1:10 to about 1:500.

The composition comprising a compound of the present invention may belyophilized. The composition comprising a compound of the presentinvention is usually presented in a vial which contains a specifiedamount of such compound.

We have found that the compounds of the present invention andcompositions of the present invention are particularly effective in thetreatment of cancer.

Thus, as described earlier, the present invention provides a method oftreating a patient in need thereof, notably a human, affected by cancerwhich comprises administering to the affected individual atherapeutically effective amount of a compound or composition accordingto the present invention. The present invention provides a compound orcomposition for use as medicament. The present invention provides acompound or composition for use in the treatment of cancer, and morepreferably a cancer selected from lung cancer, including non-small celllung cancer and small cell lung cancer, colon cancer, breast cancer,pancreas cancer, sarcoma, ovarian cancer, prostate cancer and gastriccancer.

Thus, the compounds and compositions according to the present inventionare useful for inhibiting the multiplication, or proliferation, of atumor cell or cancer cell, or for treating cancer in an animal.

The compounds and compositions according to the present invention showexcellent activity in the treatment of cancers such as lung cancerincluding non-small cell lung cancer and small cell lung cancer, coloncancer, breast cancer, pancreas cancer, sarcoma, ovarian cancer,prostate cancer and gastric cancer. Most preferred cancers are selectedfrom lung cancer including non-small cell lung cancer and small celllung cancer, breast cancer, pancreas cancer and colorectal cancer.

In the present application, by “cancer” it is meant to include tumors,neoplasias and any other malignant disease having as cause malignanttissue or cells.

The term “treating”, as used herein, unless otherwise indicated, meansreversing, attenuating, alleviating or inhibiting the progress of thedisease or condition to which such term applies, or one or more symptomsof such disorder or condition. The term “treatment”, as used herein,unless otherwise indicated, refers to the act of treating as “treating”is defined immediately above.

The compounds and compositions according to the present invention can beadministered to an animal that has also undergone surgery as treatmentfor the cancer. In one embodiment of the present invention, theadditional method of treatment is radiation therapy.

In a specific embodiment of the present invention, the compound orcomposition according to the present invention is administeredconcurrently with radiation therapy. In another specific embodiment, theradiation therapy is administered prior or subsequent to administrationof the compound or composition of the present invention, preferably atleast an hour, three hours, five hours, 12 hours, a day, a week, amonth, more preferably several months (e.g. up to three months) prior orsubsequent to administration of a compound or composition of the presentinvention.

Any radiation therapy protocol can be used depending upon the type ofcancer to be treated. For example, but not by way of limitation, x-rayradiation can be administered; in particular, high-energy megavoltage(radiation of greater than 1 MeV energy) can be used for deep tumors,and electron beam and orthovoltage x-ray radiation can be used for skincancers.

Gamma-ray emitting radioisotopes, such as radioactive isotopes ofradium, cobalt and other elements, can also be administered.

In a further embodiment of the present invention, there is provided akit comprising a therapeutically effective amount of a compoundaccording to the present invention and a pharmaceutically acceptablecarrier.

In one embodiment, the kit according to this embodiment is for use inthe treatment of cancer, and more preferably a cancer selected from lungcancer, including non-small cell lung cancer and small cell lung cancer,colon cancer, breast cancer, pancreas cancer, sarcoma, ovarian cancer,prostate cancer and gastric cancer.

In a further embodiment of the present invention, there is provided aprocess for obtaining a compound of formula I, IA, IB, IC, ID, IE, IF,IG, Ia, IAa, IBa, ICa, IDa, IEa, IFa, IGa, Ib, IAb, IBb, ICb, IDb, IEb,IFb, or IGb, or a pharmaceutically acceptable salt or ester thereof,comprising the step of reacting a compound of formula II with a compoundof formula III to give a compound of formula IV:

wherein (where allowed by possible substituent groups):

-   -   X is —NH— or —O—;    -   R₂ is a —C(═O)R^(a) group;    -   R₃ is hydrogen or a —OR^(b) group;    -   R₄ is selected from hydrogen, —CH₂OH, —CH₂OC(═O)R^(c) and        —CH₂NHProt^(NH);    -   R^(a) is selected from hydrogen, substituted or unsubstituted        C₁-C₁₂ alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl,        substituted or unsubstituted C₂-C₁₂ alkynyl;    -   R^(b) is selected from substituted or unsubstituted C₁-C₁₂        alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl, and        substituted or unsubstituted C₂-C₁₂ alkynyl;    -   R^(c) is selected from substituted or unsubstituted C₁-C₁₂        alkyl, substituted or unsubstituted    -   C₂-C₁₂ alkenyl, and substituted or unsubstituted C₂-C₁₂ alkynyl;        and    -   Prot^(NH) is a protecting group for amino;    -   with the proviso that when R₄ is hydrogen then X is —O—.

It is particularly preferred that, when R₄ is —CH₂NHProt^(NH) in thecompound of formula IV, the process further comprises the step ofdeprotecting such amino group to provide a compound of formula I, IA,IB, IC, ID, IE, IG, Ia, IAa, IBa, ICa, IDa, IEa, IGa, Ib, IAb, IBb, ICb,IDb, IEb, or IGb wherein R₄ is —CH₂NH₂ and R₁ is cyano.

In a more preferred embodiment, the process further comprises the stepof replacing the cyano group in the compound of formula IV or in thecompound of formula I, IA, IB, IC, ID, IE, IG, Ia, IAa, IBa, ICa, IDa,IEa, IGa, Ib, IAb, IBb, ICb, IDb, IEb, or IGb where R₄ is —CH₂NH₂ and R₁is cyano with a hydroxy group to give a compound of formula I, IA, IB,IC, ID, IE, IF, IG, Ia, IAa, IBa, ICa, IDa, IEa, IFa, IGa, Ib, IAb, IBb,ICb, IDb, IEb, IFb or IGb where R₁ is OH:

Preferred processes according to the present invention include:

-   -   A process that employs a compound of formula II wherein:    -   R₂ is a —C(═O)R^(a) group where R^(a) is substituted or        unsubstituted C₁-C₁₂ alkyl. Particularly preferred R^(a) is a        substituted or unsubstituted C₁-C₆ alkyl. More preferred R^(a)        is a substituted or unsubstituted alkyl group selected from        methyl, ethyl, n-propyl, isopropyl, and butyl, including        n-butyl, sec-butyl, isobutyl and tert-butyl, being methyl the        most preferred IV group.    -   A process wherein the compound of formula III is selected from a        compound of formula IIIa, IIIb and IIIc:

-   -   wherein    -   X is selected from —NH— and —O—;    -   R₃ is selected from hydrogen and OR^(b) where R^(b) is        substituted or unsubstituted C₁-C₁₂ alkyl. Particularly        preferred R^(b) is a substituted or unsubstituted C₁-C₆ alkyl.        More preferred R^(b) is a substituted or unsubstituted alkyl        group selected from methyl, ethyl, n-propyl, isopropyl, and        butyl, including n-butyl, sec-butyl, isobutyl and tert-butyl.        More preferred R³ is hydrogen or methoxy. Most preferred R³ is        hydrogen;    -   R⁴ is selected from —CH₂OH and —CH₂NHProt^(NH) where Prot^(NH)        is a protecting group for amino.    -   It is particularly preferred that the compound of formula III is        a compound of formula IIIa or IIIb.    -   A process that employs a compound of formula III, IIIa or IIIb        wherein R₄ is —CH₂OH.    -   Being preferred a process that employs a compound of formula        IIIa or IIIb wherein R₄ is as defined above.    -   Being more preferred a process that employs a compound of        formula IIIa wherein R₄ is as defined above.    -   A process that employs a compound of formula III, IIIa or IIIb        wherein R₄ is —CH₂NHProt^(NH).    -   Being preferred a process that employs a compound of formula        IIIa or IIIb wherein R₄ is as defined above.    -   Being more preferred a process that employs a compound of        formula IIIb wherein R₄ is as defined above.

EXAMPLES

Compound 1 was prepared as described in Example 20 of WO 01/87895.

Reference compounds A, B, C, D, E, F, ET-736, and PM01183 were preparedas described in WO 03/014127 (Compounds 19, 18, 44, 43, 2, 1, 26, and 27respectively).

Example 1

To a solution of 1 (0.5 g, 0.80 mmol) in acetic acid (20 mL, 0.04 M) wasadded L-tryptophanol (2-S) (533 mg, 3.0 mmol, Sigma-Aldrich). Thereaction mixture was stirred at 23° C. for 16 h and then acetic acid wasevaporated. An aqueous saturated solution of NaHCO₃ was added and themixture was extracted with CH₂Cl₂. The combined organic layers weredried over anhydrous Na₂SO₄, filtered and concentrated under vacuum.Flash chromatography (Hexane:EtOAc, 1:1) gave compounds 3-S (616 mg,97%) and 3a-S (12 mg, 2%).

3-S

R_(f)=0.50 (Hexane:EtOAc, 1:1).

¹H NMR (300 MHz, CDCl₃): δ 7.71 (s, 1H), 7.36 (dd, J=7.9, 1.0 Hz, 1H),7.27 (dd, J=8.2, 0.9 Hz, 1H), 7.13 (ddd, J=8.3, 7.0, 1.2 Hz, 1H), 7.03(ddd, J=8.0, 7.0, 1.0 Hz, 1H), 6.62 (s, 1H), 6.26 (d, J=1.4 Hz, 1H),6.04 (d, J=1.3 Hz, 1H), 5.75 (s, 1H), 5.14 (dd, J=11.7, 1.2 Hz, 1H),4.60 (s, 1H), 4.41 (s, 1H), 4.36-4.24 (m, 2H), 4.21 (d, J=2.7 Hz, 1H),3.82 (s, 3H), 3.52 (s, 1H), 3.50-3.47 (m, 1H), 3.45 (dq, J=8.4, 2.2 Hz,1H), 3.35 (t, J=10.1 Hz, 1H), 3.01-2.78 (m, 5H), 2.62 (dd, J=15.3, 4.7Hz, 1H), 2.41 (s, 1H), 2.38 (s, 3H), 2.37-2.31 (m, 1H), 2.28 (s, 3H),2.17 (s, 3H), 2.06 (s, 3H).

ESI-MS m/z: 794.2 (M+H)⁺.

3a-S

R_(f)=0.70 (Hexane:EtOAc, 1:1).

¹H NMR (500 MHz, CDCl₃): δ 7.83 (s, 1H), 7.38 (dt, J=7.9, 0.9 Hz, 1H),7.25 (dt, J=8.3, 0.9 Hz, 1H), 7.11 (ddd, J=8.2, 7.1, 1.2 Hz, 1H), 7.02(ddd, J=8.0, 7.0, 1.0 Hz, 1H), 6.62 (s, 1H), 6.24 (d, J=1.4 Hz, 1H),6.03 (d, J=1.3 Hz, 1H), 5.79 (s, 1H), 5.13 (d, J=11.7 Hz, 1H), 4.60 (s,1H), 4.39 (s, 1H), 4.36-4.22 (m, 3H), 4.17-4.09 (m, 1H), 3.91 (dd,J=10.5, 8.6 Hz, 1H), 3.83 (s, 3H), 3.51-3.41 (m, 2H), 3.04-2.92 (m, 3H),2.72 (dd, J=15.1, 4.0 Hz, 1H), 2.54-2.41 (m, 2H), 2.38 (s, 3H),2.35-2.30 (m, 1H), 2.29 (s, 3H), 2.21-2.16 (m, 1H), 2.18 (s, 3H), 2.12(s, 3H); 2.05 (s, 3H).

¹³C NMR (101 MHz, CDCl₃): δ 171.2, 170.7, 168.6, 147.5, 145.8, 143.0,141.1, 140.4, 135.6, 130.1, 129.5, 126.7, 122.2, 121.2, 120.9, 119.4,118.4, 118.2, 118.2, 113.6, 113.5, 110.9, 110.0, 109.1, 102.1, 91.4,67.2, 63.4, 61.3, 60.4, 59.7, 59.1, 54.8, 54.6, 47.7, 42.0, 41.6, 31.6,24.0, 22.6, 21.0, 15.9, 14.2, 9.7.

ESI-MS m/z: 836.2 (M+H)+.

To a solution of 3-S (616 mg, 0.77 mmol) in CH₃CN:H₂O (1.39:1, 51 mL,0.015 M) was added AgNO₃ (3.40 g, 23.3 mmol). After 3 h at 23° C., thereaction mixture was quenched with a mixture 1:1 of saturated aqueoussolutions of NaCl and NaHCO₃, stirred for 15 min, diluted with CH₂Cl₂,stirred for 5 min, and extracted with CH₂Cl₂. The combined organiclayers were dried over anhydrous Na₂SO₄, filtered, and concentratedunder vacuum. The residue obtained was purified by flash chromatography(CH₂Cl₂:CH₃OH, from 99:1 to 85:15) to give 4-S (471 mg, 78%).

R_(f)=0.50 (CH₂Cl₂:CH₃OH, 9:1).

¹H NMR (500 MHz, CDCl₃): δ 7.71 (s, 1H), 7.36 (dd, J=7.8, 1.1 Hz, 1H),7.26 (dd, J=7.8, 1.1 Hz, 1H), 7.12 (ddd, J=8.2, 7.0, 1.2 Hz, 1H), 7.03(ddd, J=8.0, 7.1, 1.0 Hz, 1H), 6.64 (s, 1H), 6.23 (d, J=1.3 Hz, 1H),6.01 (d, J=1.4 Hz, 1H), 5.75 (s, 1H), 5.25 (d, J=11.4 Hz, 1H), 4.92 (s,1H), 4.52 (br s, 3H), 4.22 (dd, J=11.4, 2.2 Hz, 1H), 4.19 (s, 1H), 3.83(s, 3H), 3.54 (br s, 2H), 3.35 (t, J=10.2 Hz, 1H), 3.26 (s, 1H),3.01-2.93 (m, 3H), 2.88 (br s, 3H), 2.63 (dd, J=15.2, 4.8 Hz, 1H), 2.38(s, 3H), 2.36-2.31 (m, 2H), 2.28 (s, 3H), 2.05 (s, 3H).

¹³C NMR (126 MHz, CDCl₃): δ 171.9, 168.6, 147.5, 145.4, 142.9, 141.2,140.7, 135.5, 130.4, 126.8, 122.3, 122.0, 121.3, 119.4, 118.4, 115.2,112.8, 111.0, 110.0, 109.6, 101.8, 81.9, 76.8, 65.2, 62.8, 62.5, 60.4,58.1, 57.9, 55.9, 55.1, 53.4, 51.6, 41.8, 41.3, 39.6, 24.1, 23.8, 20.5,15.8, 9.7.

ESI-MS m/z: 767.3 (M−H₂O+H)⁺.

(+)-HR-ESI-TOF-MS m/z 767.2788 [M−H₂O+H]⁺ (Calcd. for C₄₁H₄₃N₄O₉S:767.2745).

To a solution of 3a-S (30 mg, 0.035 mmol) in CH₃CN:H₂O (1.39:1, 2.4 mL,0.015 M) was added AgNO₃ (180 mg, 1.07 mmol). After 3 h at 23° C., thereaction mixture was quenched with a mixture 1:1 of saturated aqueoussolutions of NaCl and NaHCO₃, stirred for 15 min, diluted with CH₂Cl₂,stirred for 5 min, and extracted with CH₂Cl₂. The combined organiclayers were dried over anhydrous Na₂SO₄, filtered, and concentratedunder vacuum. The residue obtained was purified by flash chromatography(CH₂Cl₂:CH₃OH, from 99:1 to 85:15) to give 4a-S (24 mg, 83%).

R_(f)=0.60 (CH₂Cl₂:CH₃OH, 9:1).

¹H NMR (400 MHz, CDCl₃): δ 7.81 (s, 1H), 7.37 (d, J=7.8 Hz, 1H),7.30-7.21 (m, 1H), 7.06 (dddt, J=34.7, 8.0, 7.1, 1.1 Hz, 2H), 6.63 (s,1H), 6.22 (d, J=1.3 Hz, 1H), 6.02 (dd, J=12.9, 1.4 Hz, 1H), 5.74 (s,1H), 5.25-5.21 (m, 1H), 4.89 (d, J=8.7 Hz, 1H), 4.55-4.45 (m, 2H),4.30-4.18 (m, 1H), 4.14 (dd, J=10.5, 4.2 Hz, 1H), 4.00-3.88 (m, 2H),3.82 (s, 3H), 3.56-3.44 (m, 2H), 3.23 (d, J=9.0 Hz, 1H), 2.95 (d, J=15.7Hz, 2H), 2.87-2.78 (m, 2H), 2.71 (dd, J=15.0, 3.9 Hz, 1H), 2.48 (dd,J=15.1, 9.6 Hz, 1H), 2.37 (s, 3H), 2.35-2.29 (m, 1H), 2.28 (s, 3H),2.22-2.16 (m, 1H), 2.15 (s, 3H), 2.12 (s, 3H), 2.03 (s, 3H).

ESI-MS m/z: 809.2 (M−H₂O+H)⁺.

Example 2

To a solution of 1 (0.5 g, 0.80 mmol) in acetic acid (20 mL, 0.04 M) wasadded D-tryptophanol (2-R) (533 mg, 3.0 mmol, Sigma-Aldrich). Thereaction mixture was stirred at 23° C. for 16 h and then acetic acid wasevaporated. An aqueous saturated solution of NaHCO₃ was added and themixture was extracted with CH₂Cl₂. The combined organic layers weredried over anhydrous Na₂SO₄, filtered, and concentrated under vacuum.Flash chromatography (Hexane:EtOAc, 1:1) gave compound 3-R (479 mg,75%).

R_(f)=0.44 (Hexane:EtOAc, 1:1).

¹H NMR (400 MHz, CDCl₃): δ 7.61 (s, 1H), 7.39 (d, J=7.8 Hz, 1H), 7.29(d, J=9.6 Hz, 1H), 7.12 (t, J=7.3 Hz, 1H), 7.03 (t, J=7.3 Hz, 1H), 6.60(s, 1H), 6.25 (s, 1H), 6.03 (s, 1H), 5.75 (s, 1H), 5.04 (d, J=11.7 Hz,1H), 4.62 (s, 1H), 4.37 (s, 1H), 4.32-4.25 (m, 1H), 4.22 (d, J=2.7 Hz,1H), 4.19-4.09 (m, 1H), 3.82 (s, 3H), 3.77 (s, 1H), 3.64 (d, J=9.0 Hz,1H), 3.49-3.41 (m, 2H), 3.02-2.90 (m, 2H), 2.60-2.52 (m, 2H), 2.45 (d,J=14.7 Hz, 2H), 2.40 (s, 3H), 2.28 (s, 3H), 2.22-2.14 (m, 2H), 2.18 (s,3H), 2.10 (m, 3H).

ESI-MS m/z: 794.3 (M+H)⁺.

To a solution of 3-R (479 mg, 0.60 mmol) in CH₃CN:H₂O (1.39:1, 40 mL,0.015 M) was added AgNO₃ (3.03 g, 18.1 mmol). After 3 h at 23° C., thereaction mixture was quenched with a mixture 1:1 of saturated aqueoussolutions of NaCl and NaHCO₃, stirred for 15 min, diluted with CH₂Cl₂,stirred for 5 min, and extracted with CH₂Cl₂. The combined organiclayers were dried over anhydrous Na₂SO₄, filtered, and concentratedunder vacuum. The residue obtained was purified by flash chromatography(CH₂Cl₂:CH₃OH, from 99:1 to 85:15) to afford 4-R (428 mg, 91%).

R_(f)=0.45 (CH₂Cl₂:CH₃OH, 9:1).

¹H NMR (400 MHz, CDCl₃): δ 7.62 (s, 1H), 7.39 (d, J=8.1 Hz, 1H), 7.28(d, J=8.1 Hz, 1H), 7.11 (ddd, J=8.2, 7.0, 1.2 Hz, 1H), 7.02 (ddd, J=7.9,7.1, 1.0 Hz, 1H), 6.61 (s, 1H), 6.22 (d, J=1.3 Hz, 1H), 5.99 (d, J=1.3Hz, 1H), 5.73 (s, 1H), 5.17 (dd, J=11.5, 1.2 Hz, 1H), 4.86 (s, 1H),4.56-4.47 (m, 2H), 4.17 (dd, J=5.1, 1.6 Hz, 1H), 4.08 (dd, J=11.5, 2.1Hz, 1H), 3.81 (s, 3H), 3.78 (d, J=3.8 Hz, 1H), 3.64 (dd, J=10.8, 3.8 Hz,2H), 3.51 (d, J=5.1 Hz, 1H), 3.48-3.43 (m, 2H), 3.24 (d, J=8.6 Hz, 1H),3.00-2.80 (m, 2H), 2.57 (s, 1H), 2.55-2.43 (m, 1H), 2.40 (s, 3H), 2.27(s, 3H), 2.19-2.12 (m, 1H), 2.16 (s, 3H), 2.08 (s, 3H).

¹³C NMR (101 MHz, CDCl₃): δ 171.8, 168.6, 147.6, 145.4, 143.0, 141.3,140.7, 136.0, 131.1, 130.0, 129.6, 126.6, 122.1, 121.6, 121.2, 119.4,118.4, 115.6, 112.9, 111.1, 110.6, 101.8, 81.7, 65.8, 62.7, 61.8, 60.4,60.3, 57.9, 57.8, 56.1, 55.0, 52.1, 42.2, 41.3, 41.1, 23.8, 23.4, 20.5,15.7, 9.8.

ESI-MS m/z: 767.6 (M−H₂O+H)⁺.

(+)-HR-ESI-TOF-MS m/z: 767.2799 [M−H₂O+H]⁺ (Calcd. for C₄₁H₄₃N₄O₉S:767.2745).

Example 3. Synthesis of allylN—[(R)-(2-amino-3-(1H-indol-3-yl)propyl)]carbamate (9-R)

To a solution of D-tryptophanol (2-R) (2.0 g, 10.4 mmol) in CH₃CN (42mL, 4 mL/mmol) was added di-tert-butyl dicarbonate (4.6 g, 20.8 mmol).The reaction mixture was stirred at 23° C. for 3 h and concentratedunder vacuum. Flash chromatography (CH₂Cl₂:CH₃OH from 99:1 to 85:15) toafford 5-R (2.2 g, 73%).

R_(f)=0.5 (CH₂Cl₂:CH₃OH, 9:1).

¹H NMR (400 MHz, CDCl₃): δ 8.13 (s, 1H), 7.67 (dd, J=7.8, 1.1 Hz, 1H),7.38 (dd, J=8.1, 1.3 Hz, 1H), 7.29-7.10 (m, 2H), 7.06 (s, 1H), 4.82 (s,1H), 4.00 (s, 1H), 3.71 (dd, J=11.0, 3.8 Hz, 1H), 3.62 (dd, J=11.0, 5.5Hz, 1H), 3.01 (d, J=6.7 Hz, 2H), 2.14 (s, 1H), 1.44 (s, 9H).

To a solution of 5-R (2.4 g, 8.2 mmol) in CH₂Cl₂ (50 mL, 6 mL/mmol) wasadded phthalimide (2.7 g, 18.2 mmol), triphenylphosphine (4.8 g, 18.2mmol) and the mixture was cooled at 0° C. A solution of diethylazodicarboxylate solution in CH₂Cl₂ (25 mL, 3 mL/mmol) was added for 15min. The reaction was stirred at 23° C. for 16 h, concentrated undervacuum. The residue obtained was purified by flash chromatography(CH₂Cl₂:CH₃OH, from 99:1 to 85:15) to afford 6-R (3.3 g, 96%).

R_(f)=0.7 (CH₂Cl₂:CH₃OH, 9:1).

¹H NMR (400 MHz, CDCl₃): δ 8.50 (s, 1H), 7.81 (dd, J=5.5, 3.1 Hz, 2H),7.66 (dd, J=5.6, 3.2 Hz, 2H), 7.60 (d, J=7.8 Hz, 1H), 7.35 (d, J=8.0 Hz,1H), 7.19-7.04 (m, 3H), 4.81 (s, 1H), 4.40 (s, 1H), 3.83 (dd, J=13.9,3.7 Hz, 1H), 3.72 (dd, J=13.9, 9.9 Hz, 1H), 3.08-3.01 (m, 2H), 1.23 (s,9H).

To a solution of 6-R (3.25 g, 7.74 mmol) in ethanol (231 mL, 30 mL/mmol)was added hydrazine monohydrate (37 mL, 774 mmol). The reaction mixturewas stirred at 80° C. in sealed tube for 2.25 h, concentrated undervacuum. Flash chromatography (EtOAc:CH₃OH, from 100:1 to 50:50) afforded7-R (2.15 g, 96%).

R_(f)=0.2 (EtOAc:CH₃OH, 6:4).

¹H NMR (400 MHz, CD₃OD): δ 7.60 (d, J=7.9 Hz, 1H), 7.33 (d, J=8.1 Hz,1H), 7.13-7.04 (m, 2H), 7.05-6.96 (m, 1H), 4.02-3.94 (m, 1H), 2.99-2.87(m, 3H), 2.78 (dd, J=13.1, 9.7 Hz, 1H), 1.39 (s, 9H).

ESI-MS m/z: 290.2 (M+H)⁺.

To a solution of 7-R (2.15 g, 7.4 mmol) in CH₃CN (74 mL, 10 mL/mmol) andDMF (7.4 mL, 1 mL/mmol) was added N,N-diisopropylethylamine (1.06 mL,5.9 mmol) and allyl chloroformate (7.9 mL, 74 mmol). The reaction wasstirred at 23° C. for 16 h. The mixture was diluted with EtOAc, NH₄Clwas added and the mixture was extracted with EtOAc. The combined organiclayers were dried over anhydrous Na₂SO₄, filtered, and concentratedunder vacuum. The residue obtained was purified by flash chromatography(Hexane:EtOAc, from 100:1 to 1:100) to afford 8-R (1.69 g, 61%).

R_(f)=0.4 (Hexane:EtOAc, 1:1).

¹H NMR (400 MHz, CDCl₃): δ 8.25 (s, 1H), 7.62 (d, J=7.8 Hz, 1H), 7.35(dd, J=8.1, 0.9 Hz, 1H), 7.16 (dddd, J=27.8, 8.0, 7.0, 1.1 Hz, 2H), 7.04(d, J=2.4 Hz, 1H), 5.90 (ddt, J=17.3, 10.7, 5.6 Hz, 1H), 5.34-5.22 (m,1H), 5.20 (dt, J=10.5, 1.4 Hz, 1H), 5.12 (s, 1H), 4.82 (s, 1H), 4.55(dq, J=5.4, 1.7 Hz, 2H), 4.02 (s, 1H), 3.35 (dt, J=10.0, 4.7 Hz, 1H),3.21 (s, 1H), 2.95 (ddd, J=21.6, 15.4, 9.1 Hz, 2H), 1.42 (s, 9H).

ESI-MS m/z: 274.3 (M-Boc+H)⁺.

To a solution of 8-R (1.30 g, 3.50 mmol) in CH₂Cl₂ (58 mL, 16.6 mL/mmol)was added trifluoroacetic acid (30 mL, 8.3 mL/mmol). The reactionmixture was stirred at 23° C. for 1.5 h, concentrated under vacuum togive crude 9-R which was used in the next steps without furtherpurification.

R_(f)=0.2 (CH₂Cl₂:CH₃OH, 9:1).

¹H NMR (400 MHz, CDCl₃): δ 7.95 (s, 1H), 7.53 (d, J=7.8 Hz, 1H), 7.36(d, J=8.1 Hz, 1H), 7.17 (s, 1H), 7.09 (t, J=7.5 Hz, 1H), 7.03 (t, J=7.5Hz, 1H), 5.87 (ddt, J=16.4, 10.8, 5.6 Hz, 1H), 5.34-5.13 (m, 2H), 4.50(d, J=5.5 Hz, 2H), 3.62 (bs, 1H), 3.42 (dd, J=14.9, 3.9 Hz, 1H),3.36-3.20 (m, 1H), 3.11-3.00 (m, 2H).

ESI-MS m/z: 274.3 (M+H)⁺.

Example 4. Synthesis of allylN—[(S)-(2-amino-3-(1H-indol-3-yl)propyl)]carbamate (9-S)

To a solution of L-tryptophanol (2-S) (2.0 g, 10.4 mmol) in CH₃CN (42mL, 4 mL/mmol) was added Di-tert-butyl dicarbonate (4.6 g, 20.8 mmol).The reaction mixture was stirred at 23° C. for 3 h, concentrated undervacuum. Flash chromatography (CH₂Cl₂:CH₃OH, from 99:1 to 85:15) toafford 5-S (2.24 g, 73%).

R_(f)=0.5 (CH₂Cl₂:CH₃OH, 9:1).

¹H NMR (400 MHz, CDCl₃): δ 8.10 (s, 1H), 7.65 (dd, J=7.8, 1.1 Hz, 1H),7.37 (dd, J=8.1, 1.3 Hz, 1H), 7.23-7.11 (m, 2H), 7.06 (s, 1H), 4.81 (s,1H), 3.99 (s, 1H), 3.70 (dd, J=11.0, 3.8 Hz, 1H), 3.61 (dd, J=11.0, 5.5Hz, 1H), 3.00 (d, J=6.7 Hz, 2H), 2.01 (s, 1H), 1.42 (s, 9H).

To a solution of 5-S (1.2 g, 4.13 mmol) in CH₂Cl₂ (24.8 mL, 6 mL/mmol)was added phthalimide (1.33 g, 9.1 mmol), triphenylphosphine (2.4 g, 9.1mmol) and the mixture was cooled at 0° C. A solution of diethylazodicarboxylate solution (3 mL, 10.32 mmol) in CH₂Cl₂ (12.4 mL, 3mL/mmol) was added for 15 min. The reaction was stirred at 23° C. for 16h, concentrated under vacuum. The residue obtained was purified by flashchromatography (CH₂Cl₂:CH₃OH, from 99:1 to 85:15) to afford 6-S (2.8 g,>100%).

R_(f)=0.7 (CH₂Cl₂:CH₃OH, 9:1).

¹H NMR (400 MHz, CDCl₃): δ 8.49 (s, 1H), 7.80 (dd, J=5.4, 3.1 Hz, 2H),7.66 (dd, J=5.6, 3.2 Hz, 2H), 7.60 (d, J=7.8 Hz, 1H), 7.34 (d, J=8.0 Hz,1H), 7.21-7.04 (m, 3H), 4.74 (s, 1H), 4.42 (s, 1H), 3.83 (dd, J=13.9,3.7 Hz, 1H), 3.72 (dd, J=13.9, 9.9 Hz, 1H), 3.10-3.01 (m, 2H), 1.23 (s,9H).

To a solution of 6-S (0.86 g, 2.07 mmol) in ethanol (72 mL, 36 mL/mmol)was added hydrazine monohydrate (10 mL, 207 mmol). The reaction mixturewas stirred at 80° C. in sealed tube for 2.25 h, concentrated undervacuum. Flash chromatography (EtOAc:CH₃OH, from 100:1 to 50:50) toafford 7-S (1.0 g, 84%).

R_(f)=0.2 (EtOAc:CH₃OH, 6:4).

¹H NMR (400 MHz, CD₃OD): δ 7.61 (d, J=7.9 Hz, 1H), 7.35 (d, J=8.1 Hz,1H), 7.13-6.97 (m, 2H), 7.09 (s, 1H), 4.06-3.96 (m, 1H), 3.01-2.76 (m,4H), 1.38 (s, 9H).

ESI-MS m/z: 290.3 (M+H)⁺.

To a solution of 7-S (0.95 g, 3.3 mmol) in CH₃CN (33 mL, 10 mL/mmol) andDMF (3.3 mL, 1 mL/mmol) was added N,N-diisopropylethylamine (0.5 mL, 2.6mmol) and allyl chloroformate (3.5 mL, 33 mmol). The reaction wasstirred at 23° C. for 20 h. The mixture was diluted with EtOAc, NH₄Clwas added and the mixture was extracted with EtOAc. The combined organiclayers were dried over anhydrous Na₂SO₄, filtered, and concentratedunder vacuum. The residue obtained was purified by flash chromatography(Hexane:EtOAc, from 100:1 to 1:100) to afford 8-S (0.88 g, 73%).

R_(f)=0.5 (Hexane:EtOAc, 1:1).

¹H NMR (400 MHz, CDCl₃): δ 8.17 (s, 1H), 7.63 (d, J=7.8 Hz, 1H), 7.20(dd, J=8.1, 0.9 Hz, 1H), 7.13 (dddd, J=27.8, 8.0, 7.0, 1.1 Hz, 2H), 7.06(d, J=2.4 Hz, 1H), 5.90 (ddt, J=17.3, 10.7, 5.6 Hz, 1H), 5.31-5.18 (m,2H), 5.09 (s, 1H), 4.80 (s, 1H), 4.59-4.52 (m, 2H), 4.03 (s, 1H), 3.37(dt, J=10.0, 4.7 Hz, 1H), 3.21 (s, 1H), 3.05-2.87 (m, 2H), 1.42 (s, 9H).

ESI-MS m/z: 274.3 (M-Boc+H)⁺.

To a solution of 8-S (0.875 g, 2.3 mmol) in CH₂Cl₂ (38 mL, 16.6 mL/mmol)was added trifluoroacetic acid (19 mL, 8.3 mL/mmol). The reactionmixture was stirred at 23° C. for 2 h, concentrated under vacuum to givecrude 9-S which was used in the next steps without further purification.

R_(f)=0.2 (CH₂Cl₂:CH₃OH, 9:1).

¹H NMR (400 MHz, CD₃OD): δ 7.56 (d, J=7.8 Hz, 1H), 7.37 (d, J=8.1 Hz,1H), 7.21 (s, 1H), 7.13 (t, J=7.5 Hz, 1H), 7.05 (t, J=7.5 Hz, 1H), 5.94(ddt, J=16.4, 10.8, 5.6 Hz, 1H), 5.34-5.16 (m, 2H), 4.56 (d, J=5.5 Hz,2H), 3.60 (bs, 1H), 3.43 (dd, J=14.9, 3.9 Hz, 1H), 3.37-3.31 (m, 1H),3.14-2.99 (m, 2H).

ESI-MS m/z: 274.3 (M+H)⁺.

Example 5

To a solution of 1 (1.45 g, 2.33 mmol) in acetic acid (58 mL, 0.08 M)was added 9-R (0.95 g, 3.50 mmol). The reaction mixture was stirred a50° C. for 18 h and then acetic acid was evaporated. An aqueoussaturated solution of NaHCO₃ was added and the mixture was extractedwith CH₂Cl₂. The combined organic layers were dried over anhydrousNa₂SO₄. Flash chromatography (Hexane:EtOAc, 1:1) gives compound 10-R(1.3 g, 64%).

R_(f)=0.5 (Hexane:EtOAc, 1:1).

¹H NMR (400 MHz, CDCl₃): δ 7.66 (s, 1H), 7.36 (d, J=7.9 Hz, 1H), 7.27(d, J=7.9 Hz, 1H), 7.10 (ddd, J=8.3, 7.0, 1.3 Hz, 1H), 7.01 (td, J=7.5,7.0, 1.0 Hz, 1H), 6.62 (s, 1H), 6.23 (d, J=1.4 Hz, 1H), 6.01 (d, J=1.4Hz, 1H), 5.99-5.89 (m, 1H), 5.79 (s, 1H), 5.44-5.21 (m, 2H), 5.14-4.99(m, 2H), 4.63 (ddd, J=7.3, 4.4, 1.5 Hz, 2H), 4.36 (s, 1H), 4.33-4.24 (m,1H), 4.29-4.26 (m, 1H), 4.21 (d, J=2.7 Hz, 1H), 4.19-4.13 (m, 3H), 3.80(s, 3H), 3.56 (s, 1H), 3.48-3.43 (m, 3H), 3.27 (dt, J=13.2, 4.0 Hz, 1H),3.04-2.88 (m, 2H), 2.56 (dd, J=15.2, 3.8 Hz, 1H), 2.49-2.35 (m, 2H),2.31 (s, 3H), 2.28 (s, 3H), 2.17 (s, 3H), 2.07 (s, 3H).

ESI-MS m/z: 877.3 (M+H)⁺.

To a solution of 10-R (600 mg, 0.68 mmol) in CH₂Cl₂ (12 mL, 18 mL/mmol)was added bis(triphenylphosphine)palladium(II) dichloride (77 mg, 0.1mmol) and acetic acid (0.4 mL, 6.8 mmol). Tributyltin hydride (1.1 mL,4.08 mmol) was added at 0° C., the reaction mixture was stirred at 0° C.for 0.5 h and concentrated under vacuum. The crude obtained was dilutedwith EtOAc, saturated aqueous solution of NH₄Cl was added, and themixture was extracted with EtOAc. The combined organic layers were driedover anhydrous Na₂SO₄, filtered, and concentrated under vacuum. Flashchromatography (Hexane:EtOAc, from 100:1 to 1:100 and EtOAc:CH₃OH, from100:1 to 1:100) to afford 11-R (440 mg, 82%).

R_(f)=0.5 (CH₂Cl₂:CH₃OH, 1:1).

¹H NMR (400 MHz, CDCl₃): δ 7.64 (s, 1H), 7.38 (d, J=7.9 Hz, 1H), 7.29(d, J=8.1 Hz, 1H), 7.11 (ddt, J=8.3, 7.0, 1.4 Hz, 1H), 7.03 (ddt, J=8.3,7.0, 1.4 Hz, 1H), 6.58 (s, 1H), 6.24 (d, J=1.5 Hz, 1H), 6.02 (d, J=1.5Hz, 1H), 5.02 (d, J=11.8 Hz, 1H), 4.63 (s, 1H), 4.36 (s, 1H), 4.28 (d,J=5.1 Hz, 1H), 4.21 (d, J=2.2 Hz, 1H), 4.16 (s, 1H), 3.80 (s, 3H),3.51-3.39 (m, 4H), 3.32-3.13 (m, 3H), 2.95 (d, J=8.9 Hz, 2H), 2.89-2.76(m, 2H), 2.73-2.57 (m, 1H), 2.42 (d, J=14.8 Hz, 1H), 2.36 (s, 3H), 2.25(s, 3H), 2.16 (s, 3H), 2.09 (s, 3H).

ESI-MS m/z: 793.2 (M+H)⁺.

To a solution of 11-R (850 mg, 1.07 mmol) in CH₃CN:H₂O (1.39:1, 70 mL,0.015 M) was added AgNO₃ (3.64 g, 21.4 mmol). After 17 h at 23° C., thereaction was quenched with a mixture 1:1 of saturated aqueous solutionsof NaCl and NaHCO₃, stirred for 15 min, diluted with CH₂Cl₂, stirred for5 min, and extracted with CH₂Cl₂. The combined organic layers were driedover anhydrous Na₂SO₄, filtered, and concentrated under vacuum. Theresidue obtained was purified by flash chromatography (CH₂Cl₂:CH₃OH,from 99:1 to 85:15) to give 12-R (553 mg, 66%).

R_(f)=0.3 (CH₂Cl₂:CH₃OH, 9:1).

¹H NMR (500 MHz, CDCl₃): δ 7.60 (s, 1H), 7.38 (d, J=7.9 Hz, 1H), 7.28(d, J=7.9 Hz, 1H), 7.11 (ddt, J=8.3, 7.1, 1.2 Hz, 1H), 7.02 (ddt, J=8.3,7.1, 1.2 Hz, 1H), 6.58 (s, 1H), 6.22 (s, 1H), 6.00 (s, 1H), 5.16 (d,J=11.5 Hz, 1H), 4.87 (s, 1H), 4.54 (s, 1H), 4.51 (d, J=3.3 Hz, 1H), 4.17(d, J=5.4 Hz, 1H), 4.07 (dd, J=11.3, 2.2 Hz, 1H), 3.81 (s, 3H), 3.52 (d,J=5.1 Hz, 1H), 3.24 (d, J=8.8 Hz, 2H), 2.99-2.78 (m, 4H), 2.66 (dd,J=14.9, 3.5 Hz, 1H), 2.49-2.39 (m, 2H), 2.38 (s, 3H), 2.28 (m, 2H), 2.25(s, 3H), 2.21-2.16 (m, 2H), 2.15 (s, 3H), 2.08 (s, 3H).

¹³C NMR (101 MHz, CD₃OD): δ 171.7, 169.4, 148.7, 145.9, 143.7, 141.4,140.9, 136.9, 130.8, 130.0, 129.7, 126.0, 121.4, 121.0, 119.7, 119.1,118.4, 117.5, 114.9, 110.8, 107.5, 106.4, 102.1, 91.3, 63.2, 60.0, 59.0,58.6, 55.3, 54.6, 52.7, 52.4, 48.4, 45.8, 42.5, 40.2, 24.5, 23.2, 19.2,15.0, 8.2.

ESI-MS m/z: 766.2 (M−H₂O+H)⁺.

(+)-HR-ESI-TOF-MS m/z: 766.2972 [M−H₂O+H]⁺ (Calcd. for C₄₁H₄₄N₅O₈S⁺:766.2905).

To a solution of 10-R (700 mg, 0.8 mmol) in CH₃CN:H₂O (1.39:1, 87.5 mL,0.015 M) was added AgNO₃ (2.66 g, 16 mmol). After 20 h at 23° C., thereaction mixture was quenched with a mixture 1:1 of saturated aqueoussolutions of NaCl and NaHCO₃, stirred for 15 min, diluted with CH₂Cl₂,stirred for 5 min, and extracted with CH₂Cl₂. The combined organiclayers were dried over anhydrous Na₂SO₄, filtered, and concentratedunder vacuum. The residue obtained was purified by flash chromatography(CH₂Cl₂:CH₃OH, from 99:1 to 85:15) to give 13-R (438 mg, 63%).

Rf=0.40 (CH₂Cl₂:CH₃OH, 9:1).

¹H NMR (400 MHz, CDCl₃): δ 7.64 (s, 1H), 7.37 (d, J=7.9 Hz, 1H),7.32-7.20 (m, 1H), 7.11 (t, J=7.7 Hz, 1H), 7.01 (t, J=7.4 Hz, 1H), 6.62(s, 1H), 6.21 (s, 1H), 6.05-5.90 (m, 1H), 5.99 (s, 1H), 5.75 (d, J=6.0Hz, 1H), 5.40-5.07 (m, 4H), 4.88 (d, J=14.7 Hz, 1H), 4.68-4.50 (m, 3H),4.28-4.13 (m, 1H), 4.08 (dt, J=11.4, 2.4 Hz, 1H), 3.83 (s, 3H),3.68-3.40 (m, 4H), 3.37-3.19 (m, 2H), 2.98-2.79 (m, 2H), 2.59-2.36 (m,3H), 2.29 (s, 3H), 2.27 (s, 3H), 2.14 (s, 3H), 2.10-2.16 (m, 1H), 2.08(s, 3H).

ESI-MS m/z: 850.3 (M−H₂O+H)⁺.

Example 6

To a solution of 1 (955 mg, 1.5 mmol) in acetic acid (37.5 mL, 0.08 M)was added 9-S (627 mg, 2.29 mmol). The reaction mixture was stirred a50° C. for 18 h and then acetic acid was evaporated. An aqueoussaturated solution of NaHCO₃ was added and the mixture was extractedwith CH₂Cl₂. The combined organic layers were dried over anhydrousNa₂SO₄. Flash chromatography (Hexane:EtOAc, 1:1) gives compound 10-5(756 mg, 58%).

R_(f)=0.4 (Hexane:EtOAc, 1:1).

¹H NMR (400 MHz, CDCl₃): δ 7.78 (s, 1H), 7.36 (d, J=7.9 Hz, 1H), 7.24(d, J=7.9 Hz, 1H), 7.10 (ddd, J=8.3, 7.0, 1.3 Hz, 1H), 7.01 (td, J=7.5,7.0, 1.0 Hz, 1H), 6.68 (s, 1H), 6.23 (d, J=1.4 Hz, 1H), 6.01 (d, J=1.4Hz, 1H), 6.07-5.93 (m, 1H), 5.82 (s, 1H), 5.41-5.19 (m, 2H), 5.1 (d,J=11.7 Hz, 1H), 4.66 (dt, J=5.9, 1.3 Hz, 1H), 4.57 (s, 1H), 4.37 (s,1H), 4.33-4.20 (m, 3H), 3.81 (s, 3H), 3.46 (d, J=4.2 Hz, 2H), 3.22-3.13(m, 1H), 3.11-2.88 (m, 4H), 2.66 (dd, J=15.2, 4.2 Hz, 1H), 2.51 (dd,J=15.3, 6.0 Hz, 1H), 2.43-2.32 (m, 2H), 2.31 (s, 3H), 2.26 (s, 3H), 2.19(s, 3H), 2.04 (s, 3H).

ESI-MS m/z: 877.3 (M+H)⁺.

To a solution of 10-S (650 mg, 0.72 mmol) in CH₂Cl₂ (13.3 mL, 18mL/mmol) was added bis(triphenylphosphine)palladium(II) dichloride (83mg, 0.11 mmol) and acetic acid (0.42 mL, 7.4 mmol). Tributyltin hydride(1.2 mL, 4.4 mmol) was added at 0° C., the reaction mixture was stirredat 23° C. for 0.5 h, and concentrated under vacuum. Flash chromatography(Hexane:EtOAc, from 100:1 to 1:100 and EtOAc:CH₃OH, from 100:1 to 1:100)to afford 11-S (445 mg, 78%).

R_(f)=0.5 (CH₂Cl₂:CH₃OH, 1:1).

¹H NMR (400 MHz, CDCl₃): δ 7.74 (s, 1H), 7.36 (d, J=7.9 Hz, 1H), 7.26(d, J=8.1 Hz, 1H), 7.12 (ddt, J=8.3, 7.0, 1.4 Hz, 1H), 7.02 (ddt, J=8.3,7.0, 1.4 Hz, 1H), 6.62 (s, 1H), 6.26 (d, J=1.5 Hz, 1H), 6.04 (d, J=1.5Hz, 1H), 5.12 (d, J=11.8 Hz, 1H), 4.59 (s, 1H), 4.42 (s, 1H), 4.36-4.17(m, 3H), 3.81 (s, 3H), 3.51-3.39 (m, 3H), 2.98-2.75 (m, 4H), 2.69-2.60(m, 2H), 2.47 (d, J=16.1 Hz, 1H), 2.38 (s, 3H), 2.35-2.17 (m, 2H), 2.28(s, 3H), 2.13 (s, 3H), 2.04 (s, 3H).

ESI-MS m/z: 793.3 (M+H)⁺.

To a solution of 11-5 (435 mg, 0.55 mmol) in CH₃CN:H₂O (1.39:1, 38.5 mL,0.015 M) was added AgNO₃ (1.84 g, 11 mmol). After 24 h at 23° C., thereaction was quenched with a mixture 1:1 of saturated aqueous solutionsof NaCl and NaHCO₃, stirred for 15 min, diluted with CH₂Cl₂, stirred for5 min, and extracted with CH₂Cl₂. The combined organic layers were driedover anhydrous Na₂SO₄, filtered, and concentrated under vacuum. Theresidue obtained was purified by flash chromatography (CH₂Cl₂:CH₃OH,from 99:1 to 85:15) to give 12-S (152 mg, 35%).

R=0.2 (CH₂Cl₂:CH₃OH, 9:1).

¹H NMR (500 MHz, CD₃OD): δ 7.34 (dd, J=7.7, 1.5 Hz, 1H), 7.28 (dd,J=7.7, 1.5 Hz, 1H), 7.04 (ddt, J=8.2, 7.0, 1.1 Hz, 1H), 6.95 (ddt,J=8.2, 7.0, 1.2 Hz, 1H), 6.55 (s, 1H), 6.31-6.25 (m, 1H), 6.15-6.05 (m,1H), 5.31 (d, J=11.4 Hz, 1H), 4.91 (s, 1H), 4.64 (s, 1H), 4.40-4.19 (m,3H), 3.76 (s, 3H), 3.64 (d, J=5.2 Hz, 1H), 3.44 (d, J=9.0 Hz, 1H),3.03-2.85 (m, 4H), 2.85-2.65 (m, 2H), 2.59 (d, J=15.6 Hz, 1H), 2.52-2.39(m, 2H), 2.37 (s, 3H), 2.27 (s, 3H), 2.09 (s, 3H), 2.00 (s, 3H).

¹³C NMR (126 MHz, CD₃OD): δ 171.4, 169.3, 148.6, 145.8, 143.5, 141.2,140.8, 136.5, 131.2, 130.3, 129.5, 126.3, 121.6, 121.2, 119.8, 119.4,118.6, 117.5, 114.9, 111.0, 107.5, 107.4, 102.2, 91.1, 63.5, 60.5, 59.2,58.5, 55.3, 54.7, 53.4, 52.7, 48.6, 44.7, 42.7, 39.9, 24.3, 23.4, 19.2,15.1, 8.2.

ESI-MS m/z: 766.2 (M−H₂O+H)⁺.

(+)-HR-ESI-TOF-MS m/z: 766.2958 [M−H₂O+H]⁺ (Calcd. for C₄₁H₄₄N₅O₈S:766.2905).

To a solution of 10-S (5 mg, 0.006 mmol) in CH₃CN:H₂O (1.39:1, 0.5 mL,0.015 M) was added AgNO₃ (29 mg, 0.17 mmol). After 20 h at 23° C., thereaction mixture was quenched with a mixture 1:1 of saturated aqueoussolutions of NaCl and NaHCO₃, stirred for 15 min, diluted with CH₂Cl₂,stirred for 5 min, and extracted with CH₂Cl₂. The combined organiclayers were dried over anhydrous Na₂SO₄, filtered, and concentratedunder vacuum. The residue obtained was purified by flash chromatography(CH₂Cl₂:CH₃OH, from 99:1 to 85:15) to give 13-S (5 mg, 100%).

R_(f)=0.40 (CH₂Cl₂:CH₃OH, 9:1).

¹H NMR (400 MHz, CDCl₃): δ 7.75 (s, 1H), 7.37 (d, J=7.9 Hz, 1H),7.32-7.20 (m, 1H), 7.12 (t, J=7.7 Hz, 1H), 7.02 (t, J=7.4 Hz, 1H), 6.84(s, 1H), 6.24 (s, 1H), 6.08-5.97 (m, 1H), 6.01 (s, 1H), 5.87 (s, 1H),5.42-5.19 (m, 4H), 4.88 (s, 1H), 4.69-4.65 (m, 2H), 4.58 (s, 1H),4.28-4.13 (m, 2H), 3.84 (s, 3H), 3.68-3.40 (m, 2H), 3.24-3.15 (m, 2H),3.08-2.90 (m, 2H), 2.73-2.57 (m, 2H), 2.53-2.37 (m, 3H), 2.34 (s, 3H),2.25 (s, 3H), 2.14 (s, 3H), 2.10-2.16 (m, 1H), 2.03 (s, 3H).

ESI-MS m/z: 850.3 (M−H₂O+H)⁺.

Example 7. Synthesis of Reference Compounds 14-S and 15-S

To a solution of 1 (50 mg, 0.08 mmol) in acetic acid (1 mL, 0.08 M) wasadded L-tryptophan (50 mg, 0.24 mmol). The reaction mixture was stirredat 50° C. for 17 h and then acetic acid was evaporated. An aqueoussaturated solution of NaHCO₃ was added and the mixture was extractedwith CH₂Cl₂. The combined organic layers were dried over anhydrousNa₂SO₄, filtered and concentrated under vacuum. Flash chromatography(CH₂Cl₂:CH₃OH, from 99:1 to 80:20) gave compound 14-5 (58 mg, 90%).

R_(f)=0.20 (CH₂Cl₂:CH₃OH, 10:1).

¹H NMR (400 MHz, CDCl₃): δ 7.77 (s, 1H), 7.39 (d, J=7.9 Hz, 1H), 7.25(d, J=7.9 Hz, 1H), 7.13 (ddd, J=8.2, 7.0, 1.2 Hz, 1H), 7.04 (td, J=7.5,7.1, 1.0 Hz, 1H), 6.56 (s, 1H), 6.24 (d, J=1.3 Hz, 1H), 6.03 (d, J=1.3Hz, 1H), 5.15 (d, J=11.7 Hz, 1H), 4.62 (s, 1H), 4.43 (s, 1H), 4.35 (dd,J=11.7, 2.1 Hz, 1H), 4.28 (dd, J=5.2, 1.6 Hz, 1H), 4.20 (s, 1H), 3.78(s, 3H), 3.52-3.41 (m, 4H), 3.07-2.88 (m, 2H), 2.91-2.80 (m, 2H),2.42-2.21 (m, 2H), 2.35 (s, 3H), 2.27 (s, 3H), 2.14 (s, 3H), 2.04 (s,3H).

ESI-MS m/z: 808.6 (M+H)⁺.

To a solution of 14-S (52 mg, 0.066 mmol) in CH₃CN:H₂O (2:1, 4.5 mL,0.015 M) was added AgNO₃ (164 mg, 1.45 mmol). After 20 h at 23° C., amixture 1:1 of saturated aqueous solutions of NaCl and NaHCO₃ was added,stirred for 15 min, diluted with CH₂Cl₂, stirred for 30 min, andextracted with CH₂Cl₂. The combined organic layers were dried overanhydrous Na₂SO₄, filtered and concentrated under vacuum. The residueobtained was purified by flash chromatography (CH₂Cl₂:CH₃OH, from 99:1to 70:30) to afford 15-S (18 mg, 35%).

R_(f)=0.15 (CH₂Cl₂:CH₃OH, 9:1).

¹H NMR (400 MHz, CD₃OD): δ 7.76 (s, 1H), 7.40 (d, J=7.8 Hz, 1H), 7.25(d, J=7.8 Hz, 1H), 7.14 (t, J=7.4 Hz, 1H), 7.04 (t, J=7.4 Hz, 1H), 6.58(s, 1H), 6.23 (d, J=1.3 Hz, 1H), 6.01 (d, J=1.3 Hz, 1H), 5.28 (d, J=12.7Hz, 1H), 4.95 (s, 1H), 4.53 (s, 1H), 4.28 (dd, J=11.4, 2.0 Hz, 1H), 4.21(s, 1H), 3.80 (s, 3H), 3.58 (s, 1H), 3.52-3.47 (m, 2H), 3.28 (s, 1H),3.03 (dd, J=15.8, 5.2 Hz, 1H), 2.91-2.82 (m, 3H), 2.44 (d, J=15.4 Hz,1H), 2.36 (s, 3H), 2.35-2.31 (m, 1H), 2.28 (s, 3H), 2.15 (s, 3H), 2.03(s, 3H).

¹³C NMR (101 MHz, CDCl₃): δ 173.7, 171.2, 168.7, 147.5, 145.7, 142.8,141.2, 140.8, 135.6, 129.8, 126.3, 122.8, 121.5, 121.2, 119.9, 118.6,117.7, 115.0, 111.1, 101.9, 81.5, 66.8, 62.9, 60.4, 57.9, 55.8, 55.1,52.3, 42.3, 41.3, 38.3, 31.9, 29.4, 28.9, 24.5, 24.0, 23.8, 22.7, 20.5,16.0, 9.7.

ESI-MS m/z: 781.6 (M−H₂O+H)⁺.

(+)-HR-ESI-TOF-MS m/z: 781.2610 [M−H₂O+H]⁺ (Calcd. for C₄₁H₄₁N₄O₁₀S:781.2538).

Example 8 A) Synthesis of (S)-5-methoxy-tryptophanol (17-S)

To a solution of LiAlH₄ (23.4 mL, 1.0 M in THF, 23.4 mmol) at −40° C.was added carefully H₂SO₄ (0.31 mL, 5.57 mmol) and a suspension of5-methoxy-L-tryptophan (16-S) (1.0 g, 4.26 mmol, Chem-Impex) in THF(13.4 mL, 0.3 M). The reaction mixture was left evolution at 23° C.,heated for 3 h at 80° C. and 18 h at 23° C. Cool at −21° C. the reactionmixture was quenched carefully with NaOH 2N until basic pH. EtOAc wasadded and the mixture filtered through Celite® and washed with CH₃OH.The crude was concentrated under vacuum to give 17-S as a crude whichwas used in the next step without further purification.

R_(f)=0.2 (CH₂Cl₂:CH₃OH, 4:1).

¹H NMR (400 MHz, CDCl₃): δ 7.19 (dt, J=8.8, 0.7 Hz, 1H), 7.06-7.00 (m,2H), 6.72 (dd, J=8.8, 2.4 Hz, 1H), 3.77 (s, 3H), 3.63-3.48 (m, 1H),3.42-3.33 (m, 1H), 3.17-3.06 (m, 1H), 2.86 (ddt, J=14.3, 6.1, 0.8 Hz,1H), 2.66 (dd, J=14.3, 7.5 Hz, 1H).

ESI-MS m/z: 221.4 (M+H)⁺.

B) Synthesis of (R)-5-methoxy-tryptophanol (17-R)

To a solution of LiAlH₄ (11.7 mL, 1.0 M in THF, 11.7 mmol) at −40° C.was added carefully H₂SO₄ (0.31 mL, 5.75 mmol) and a suspension of5-methoxy-D-tryptophan (16-R) (0.5 g, 2.13 mmol, Aldrich) in THF (6.7mL, 0.3 M). The reaction mixture was left evolution at 23° C., heatedfor 3.5 h at 80° C. and 18 h at 23° C. Cool at −21° C. the reactionmixture was quenched carefully with NaOH 2N until basic pH. EtOAc wasadded and the mixture filtered through Celite® and washed with CH₃OH.The crude was concentrated under vacuum to give 17-R as a crude whichwas used in the next step without further purification.

R_(f)=0.2 (CH₂Cl₂:CH₃OH, 4:1).

¹H NMR (400 MHz, CD₃OD): δ 7.20 (d, J=8.9 Hz, 1H), 7.06-6.96 (m, 2H),6.71 (dd, J=8.8, 2.5 Hz, 1H), 3.75 (s, 3H), 3.62-3.52 (m, 1H), 3.37 (dd,J=10.8, 7.0 Hz, 1H), 3.09 (br s, 1H), 2.82 (dd, J=14.3, 5.9 Hz, 1H),2.62 (dd, J=14.4, 7.6 Hz, 1H).

ESI-MS m/z: 221.6 (M+H)⁺.

Example 9

To a solution of 1 (530 mg, 0.85 mmol) in acetic acid (10.6 mL, 0.08 M)was added 17-S (469 mg, 2.13 mmol). The reaction mixture was stirred at50° C. for 18 h and then acetic acid was evaporated. An aqueoussaturated solution of NaHCO₃ was added and the mixture was extractedwith CH₂Cl₂. The combined organic layers were dried over anhydrousNa₂SO₄, filtered, and concentrated under vacuum. Flash chromatography(Hexane:EtOAc, 1:1) gave compound 18-S (420 mg, 60%).

R_(f)=0.3 (Hexane:EtOAc, 1:1).

¹H NMR (400 MHz, CD₃OD): δ 7.13 (d, J=8.8 Hz, 1H), 6.80 (d, J=2.4 Hz,1H), 6.66 (dd, J=8.8, 2.5 Hz, 1H), 6.51 (s, 1H), 6.27 (s, 1H), 6.11 (s,1H), 5.21 (d, J=11.7 Hz, 1H), 4.67 (s, 1H), 4.49-4.29 (m, 4H), 3.75 (s,3H), 3.73 (s, 3H), 3.47 (t, J=5.8 Hz, 3H), 3.37 (d, J=5.1 Hz, 1H),3.01-2.81 (m, 2H), 2.75 (d, J=7.4 Hz, 1H), 2.66 (dd, J=15.1, 4.1 Hz,1H), 2.55-2.35 (m, 4H), 2.34 (s, 3H), 2.28 (s, 3H), 2.11 (s, 3H), 1.99(s, 3H).

ESI-MS m/z: 824.3 (M+H)⁺.

To a solution of 18-S (420 mg, 0.519 mmol) in CH₃CN:H₂O (1.39:1, 36 mL,0.015 M) was added AgNO₃ (2.60 g, 15.3 mmol). After 3 h at 23° C., thereaction mixture was quenched with a mixture 1:1 of saturated aqueoussolutions of NaCl and NaHCO₃, stirred for 15 min, diluted with CH₂Cl₂,stirred for 5 min, and extracted with CH₂Cl₂. The combined organiclayers were dried over anhydrous Na₂SO₄, filtered, and concentratedunder vacuum. The residue obtained was purified by flash chromatography(CH₂Cl₂:CH₃OH, from 99:1 to 85:15) to obtain 19-S (250 mg, 60%).

R_(f)=0.45 (CH₂Cl₂:CH₃OH, 9:1).

¹H NMR (500 MHz, CD₃OD): δ 7.15 (dd, J=8.9, 0.6 Hz, 1H), 6.82 (dd,J=2.4, 0.6 Hz, 1H), 6.68 (dd, J=8.8, 2.5 Hz, 1H), 6.54 (s, 1H), 6.27 (d,J=1.3 Hz, 1H), 6.08 (d, J=1.3 Hz, 1H), 5.30 (d, J=11.5 Hz, 1H), 4.62 (s,1H), 4.34 (dd, J=11.4, 2.0 Hz, 1H), 4.31-4.27 (m, 2H), 3.76 (s, 3H),3.75 (s, 3H), 3.66-3.58 (m, 1H), 3.55-3.45 (m, 2H), 3.42 (d, J=7.8 Hz,1H), 2.93-2.73 (m, 3H), 2.68 (dd, J=15.1, 4.2 Hz, 1H), 2.54 (d, J=15.4Hz, 1H), 2.42 (dd, J=15.1, 10.1 Hz, 2H), 2.35 (s, 3H), 2.29 (s, 3H),2.09 (s, 3H), 2.00 (s, 3H).

¹³C NMR (126 MHz, CD₃OD): δ 172.7, 170.8, 155.1, 149.9, 147.2, 145.0,142.6, 142.2, 133.1, 132.4, 132.1, 131.3, 128.1, 122.5, 121.6, 120.3,116.4, 113.0, 112.9, 111.4, 109.0, 103.6, 100.8, 92.5, 66.6, 65.0, 61.7,60.4, 59.9, 56.7, 56.1, 54.8, 54.1, 51.7, 44.1, 41.3, 30.7, 25.4, 24.7,20.6, 16.3, 9.5.

ESI-MS m/z: 798.1 (M−H₂O+H)⁺.

(+)-HR-ESI-TOF-MS m/z: 797.2899 [M−H₂O+H]⁺ (Calcd. for C₄₂H₄₅N₄O₁₀S797.2851).

Example 10

To a solution of 1 (311 mg, 0.50 mmol) in acetic acid (6.25 mL, 0.08 M)was added 17-R (220 mg, 1.0 mmol). The reaction mixture was stirred at50° C. for 18 h and then acetic acid was evaporated. An aqueoussaturated solution of NaHCO₃ was added and the mixture was extractedwith CH₂Cl₂. The combined organic layers were dried over anhydrousNa₂SO₄, filtered, and concentrated under vacuum. Flash chromatography(Hexane:EtOAc, 1:1) gave compound 18-R (280 mg, 68%).

R_(f)=0.3 (Hexane:EtOAc, 1:1).

¹H NMR (400 MHz, CDCl₃): δ 7.53 (s, 1H), 7.18 (d, J=8.7 Hz, 1H), 6.82(d, J=2.4 Hz, 1H), 6.78 (dd, J=8.6, 2.3 Hz, 1H), 6.60 (s, 1H), 6.23 (s,1H), 6.02 (s, 1H), 5.76 (s, 1H), 5.04 (d, J=11.7 Hz, 1H), 4.62 (s, 1H),4.36 (s, 1H), 4.28 (d, J=5.0 Hz, 1H), 4.24-4.09 (m, 3H), 3.81 (s, 3H),3.79 (s, 3H), 3.64 (s, 1H), 3.47-3.40 (m, 3H), 3.01-2.90 (m, 2H), 2.53(d, J=6.9 Hz, 2H), 2.45-2.41 (m, 1H), 2.40 (s, 3H), 2.27 (s, 3H),2.22-2.14 (m, 1H), 2.18 (s, 3H), 2.06 (s, 3H).

ESI-MS m/z: 824.3 (M+H)⁺.

To a solution of 18-R (330 mg, 0.40 mmol) in CH₃CN:H₂O (1.39:1, 28 mL,0.015 M) was added AgNO₃ (2.04 g, 12.0 mmol). After 3 h at 23° C., thereaction was quenched with a mixture 1:1 of saturated aqueous solutionsof NaCl and NaHCO₃, stirred for 15 min, diluted with CH₂Cl₂, stirred for5 min, and extracted with CH₂Cl₂. The combined organic layers were driedover anhydrous Na₂SO₄, filtered, and concentrated under vacuum. Theresidue obtained was purified by flash chromatography (CH₂Cl₂:CH₃OH,from 99:1 to 85:15) to obtain 19-R (224 mg, 69%).

R_(f)=0.44 (CH₂Cl₂:CH₃OH, 9:1).

¹H NMR (500 MHz, CD₃OD): δ 7.14 (dd, J=8.8, 0.5 Hz, 1H), 6.83 (d, J=2.5Hz, 1H), 6.68 (dd, J=8.8, 2.5 Hz, 1H), 6.59 (s, 1H), 6.26 (d, J=1.4 Hz,1H), 6.07 (d, J=1.4 Hz, 1H), 5.21 (d, J=11.5 Hz, 1H), 4.68-4.55 (m, 1H),4.32-4.25 (m, 2H), 4.12 (dd, J=11.5, 2.1 Hz, 1H), 3.75 (s, 3H), 3.74 (s,3H), 3.60 (d, J=5.2 Hz, 1H), 3.57-3.45 (m, 3H), 3.41 (d, J=8.8 Hz, 1H),2.97-2.83 (m, 3H), 2.73 (dd, J=15.0, 3.4 Hz, 1H), 2.69 (d, J=14.9 Hz,1H), 2.34 (s, 3H), 2.30 (s, 3H), 2.20 (dd, J=15.1, 10.4 Hz, 1H), 2.12(s, 3H), 2.11-2.08 (m, 1H), 2.05 (s, 3H).

¹³C NMR (126 MHz, CD₃OD): δ 173.0, 170.8, 155.0, 149.8, 147.3, 145.0,142.8, 142.3, 133.5, 133.1, 132.2, 132.1, 131.1, 130.5, 127.8, 122.5,121.7, 120.0, 116.4, 113.5, 112.9, 111.4, 110.2, 103.5, 100.9, 92.6,66.8, 64.5, 61.3, 60.4, 60.0, 56.8, 56.1, 55.9, 54.1, 44.1, 41.3, 25.6,24.5, 20.6, 16.2, 9.6.

ESI-MS m/z: 797.4 (M−H₂O+H)⁺.

(+)-HR-ESI-TOF-MS m/z: 797.2896 [M−H₂O+H]⁺ (Calcd. for C₄₂H₄₅N₄O₁₀S797.2851).

Example 11. Synthesis of allylN—[(S)-2-amino-3-(5-methoxy-1H-indol-3-yl)propyl)]carbamate (24-S)

To a solution of 17-S (6.9 g, 31.4 mmol) in CH₃CN (126 mL, 4 mL/mmol)was added di-tert-butyl dicarbonate (13.7 g, 62.8 mmol). The reactionmixture was stirred at 23° C. for 5.5 h, concentrated under vacuum.Flash chromatography (CH₂Cl₂:CH₃OH, from 99:1 to 85:15) gives 20-S (4.5g, 45%).

R_(f)=0.6 (CH₂Cl₂:CH₃OH, 9:1).

¹H NMR (400 MHz, CDCl₃): δ 8.04 (s, 1H), 7.25 (d, J=8.4 Hz, 1H), 7.10(d, J=2.4 Hz, 1H), 7.03 (s, 1H), 6.87 (dd, J=8.8, 2.5 Hz, 1H), 4.83 (s,1H), 3.98 (s, 1H), 3.87 (s, 3H), 3.73-3.58 (m, 2H), 2.96 (d, J=6.6 Hz,2H), 1.42 (s, 9H).

To a solution of 20-S (4.5 g, 14 mmol) in CH₂Cl₂ (84 mL, 6 mL/mmol) wasadded phthalimide (4.5 g, 30.9 mmol), triphenylphosphine (8.1 g, 30.9mmol) and the mixture was cooled at 0° C. A solution of 40% of diethylazodicarboxylate in CH₂Cl₂ (10.4 mL, 35 mmol) was added for 15 min. Thereaction was stirred at 23° C. for 18 h, concentrated under vacuum. Theresidue obtained was purified by flash chromatography (Hexane:EtOAc,from 99:1 to 85:15) to yield 21-5 (5.8 g, 92%).

R_(f)=0.55 (Hexane:EtOAc, 1:1).

¹H NMR (400 MHz, CDCl₃): δ 8.48 (s, 1H), 7.78 (dd, J=5.5, 3.1 Hz, 2H),7.69-7.61 (m, 2H), 7.21 (d, J=8.8 Hz, 1H), 7.06 (dd, J=18.5, 2.4 Hz,2H), 6.81 (dd, J=8.8, 2.4 Hz, 1H), 4.87 (s, 1H); 4.39 (s, 1H), 3.87 (s,3H), 3.83-3.66 (m, 2H), 2.98 (d, J=6.1 Hz, 2H), 1.20 (s, 9H).

To a solution of 21-S (6.29 g, 14 mmol) in ethanol (420 mL, 30 mL/mmol)was added hydrazine monohydrate (61.1 mL, 1260 mmol). The reactionmixture was stirred at 80° C. in sealed tube for 2 h, concentrated undervacuum. Flash chromatography (CH₂Cl₂:CH₃OH, from 100:1 to 50:50) affords22-S (4.2 g, 95%).

R_(f)=0.1 (CH₂Cl₂:CH₃OH, 8:2).

¹H NMR (400 MHz, CDCl₃): δ 7.22 (d, J=8.8 Hz, 1H), 7.12 (d, J=2.4 Hz,1H), 7.06 (s, 1H), 6.76 (dd, J=8.8, 2.4 Hz, 1H), 4.06-3.97 (m, 1H), 3.82(s, 3H), 3.06-2.82 (m, 4H), 1.37 (s, 9H).

To a solution of 22-S (4.0 g, 12.52 mmol) in CH₃CN (125 mL, 10 mL/mmol)and DMF (12 mL, 1 mL/mmol) was added N,N-diisopropylethylamine (1.8 mL,10 mmol) and allyl chloroformate (13.3 mL, 125 mmol). The reaction wasstirred at 23° C. for 5 h. The mixture was diluted with EtOAc and NH₄Clwas added and the mixture was extracted with EtOAc. The combined organiclayers were dried over anhydrous Na₂SO₄, filtered, and concentratedunder vacuum. The residue obtained was purified by flash chromatography(Hexane:EtOAc, from 100:1 to 1:100) to obtain 23-S (2.65 g, 52%).

R_(f)=0.5 (Hexane:EtOAc, 1:1).

¹H NMR (400 MHz, CDCl₃): δ 8.11 (s, 1H), 7.28-7.20 (m, 1H), 7.04 (d,J=13.1 Hz, 2H), 6.85 (dd, J=8.9, 2.4 Hz, 1H), 5.97-5.82 (m, 1H),5.33-5.24 (m, 1H), 5.19 (dt, J=10.4, 1.3 Hz, 1H), 5.11 (s, 1H), 4.82 (s,1H), 4.55 (d, J=5.6 Hz, 2H), 4.01 (s, 1H), 3.86 (s, 3H), 3.37 (d, J=13.7Hz, 1H), 3.21 (s, 1H), 2.89 (dd, J=14.5, 7.0 Hz, 1H), 1.41 (s, 9H).

To a solution of 23-S (2.60 g, 6.44 mmol) in CH₂Cl₂ (106 mL, 16.6mL/mmol) was added trifluoroacetic acid (54 mL, 8.3 mL/mmol). Thereaction mixture was stirred at 23° C. for 1.5 h, concentrated undervacuum to afford 24-S (3.9 g, 100%).

R_(f)=0.1 (CH₂Cl₂:CH₃OH, 9:1).

¹H NMR (400 MHz, CD₃OD): δ 8.27 (s, 1H), 7.25 (dd, J=9.0, 2.4 Hz, 1H),7.10 (s, 1H), 6.96 (d, J=2.3 Hz, 1H), 6.87 (dd, J=9.0, 2.4 Hz, 1H), 5.81(ddt, J=16.3, 10.9, 5.7 Hz, 1H), 5.23 (dd, J=19.3, 13.6 Hz, 2H), 4.49(d, J=5.9 Hz, 2H), 3.82 (s, 3H), 3.81-3.55 (m, 1H), 3.62-3.39 (m, 2H),3.08 (qd, J=15.1, 7.3 Hz, 2H).

Example 12

To a solution of 1 (120 mg, 0.19 mmol) in acetic acid (6 mL, 0.08 M) wasadded 24-S (117 mg, 0.35 mmol). The reaction mixture was stirred at 23°C. for 18 h and then acetic acid was evaporated. An aqueous saturatedsolution of NaHCO₃ was added and the mixture was extracted with CH₂Cl₂.The combined organic layers were dried over anhydrous Na₂SO₄, filtered,and concentrated under vacuum. Flash chromatography (Hexane:EtOAc, 1:1)gives compound 25-S (95 mg, 54%).

R_(f)=0.4 (Hexane:EtOAc, 1:1).

¹H NMR (400 MHz, CDCl₃): δ 7.64 (s, 1H), 7.14 (d, J=8.8 Hz, 1H), 6.80(s, 1H), 6.77 (d, J=8.8 Hz, 1H), 6.68 (s, 1H), 6.24 (s, 1H), 6.03 (s,1H), 6.02-5.93 (m, 1H), 5.76 (s, 1H), 5.38 (d, J=10.5 Hz, 1H), 5.26 (d,J=10.5 Hz, 1H), 5.11 (d, J=11.7 Hz, 1H), 4.66 (d, J=5.6 Hz, 2H), 4.57(s, 1H), 4.37 (s, 1H), 4.33-4.19 (m, 3H), 3.82 (s, 3H), 3.79 (s, 3H),3.46 (s, 2H), 3.17 (s, 1H), 3.10-2.90 (m, 3H), 2.68-2.45 (m, 2H),2.38-2.33 (m, 1H), 2.32 (s, 3H), 2.27 (s, 3H), 2.16 (s, 3H), 2.04 (s,2H).

ESI-MS m/z: 907.1 (M+H)⁺.

To a solution of 25-S (90 mg, 0.1 mmol) in CH₂Cl₂ (2 mL, 18 mL/mmol) wasadded bis(triphenylphosphine)palladium(II)dichloride (12 mg, 0.1 mmol)and acetic acid (0.056 mL, 0.99 mmol). Tributyltin hydride (0.16 mL,0.60 mmol) was added at 0° C., the reaction mixture was stirred at 0° C.for 0.5 h, and concentrated under vacuum. Flash chromatography(Hexane:EtOAc, from 100:1 to 1:100 and EtOAc:CH₃OH, from 100:1 to 1:100)to afford 26-S (75 mg, 92%).

R_(f)=0.25 (CH₂Cl₂:CH₃OH, 1:1).

¹H NMR (400 MHz, CDCl₃): δ 7.62 (s, 1H), 7.15 (d, J=9.3 Hz, 1H),6.81-6.76 (m, 2H), 6.72 (s, 1H), 6.25 (d, J=1.2 Hz, 1H), 6.03 (d, J=1.2Hz, 1H), 5.12 (d, J=11.7 Hz, 1H), 4.57 (s, 1H), 4.41 (s, 1H), 4.36-4.24(m, 2H), 4.20 (d, J=11.7 Hz, 1H), 3.82 (s, 3H), 3.79 (s, 3H), 3.44 (dd,J=22.0, 7.1 Hz, 2H), 3.08-2.78 (m, 4H), 2.73-2.64 (m, 2H), 2.41-2.22 (m,3H), 2.28 (s, 3H), 2.25-2.15 (m, 1H), 2.14 (s, 3H), 2.08 (s, 3H), 2.04(s, 3H).

ESI-MS m/z: 823.3 (M+H)⁺.

To a solution of 26-S (70 mg, 0.085 mmol) in CH₃CN:H₂O (1.39:1, 6 mL,0.015 M) was added AgNO₃ (335 mg, 1.7 mmol). After 18 h at 23° C., thereaction was quenched with a mixture 1:1 of saturated aqueous solutionsof NaCl and NaHCO₃, stirred for 15 min, diluted with CH₂Cl₂, stirred for5 min, and extracted with CH₂Cl₂. The combined organic layers were driedover anhydrous Na₂SO₄, filtered, and concentrated under vacuum. Theresidue obtained was purified by flash chromatography (CH₂Cl₂:CH₃OH,from 99:1 to 85:15) to give 27-S (23 mg, 33%).

R_(f)=0.2 (CH₂Cl₂:CH₃OH, 9:1).

¹H NMR (400 MHz, CDCl₃): δ 7.62 (s, 1H), 7.15 (d, J=7.8 Hz, 1H), 6.78(s, 1H), 6.75 (d, J=7.8 Hz, 1H), 6.21 (d, J=1.5 Hz, 1H), 6.01 (d, J=1.5Hz, 1H), 5.78 (s, 1H), 5.22 (d, J=11.5 Hz, 1H), 4.90 (s, 1H), 4.58-4.42(m, 3H), 4.29-4.10 (m, 2H), 3.84-3.80 (m, 1H), 3.83 (s, 3H), 3.79 (s,3H), 3.53-3.48 (m, 2H), 3.22 (d, J=8.7 Hz, 1H), 3.12 (s, 1H), 3.02 (d,J=12.8 Hz, 1H), 2.89-2.64 (m, 3H), 2.46 (s, 3H), 2.42-2.34 (m, 2H), 2.27(s, 3H), 2.12 (s, 3H), 2.03 (s, 3H).

¹³C NMR (126 MHz, CDCl₃): δ 172.1, 168.7, 154.0, 147.6, 145.6, 143.0,141.2, 140.8, 131.6, 130.6, 129.6, 127.1, 121.8, 120.9, 118.4, 115.2,112.5, 111.8, 101.8, 100.2, 81.5, 62.6, 60.6, 58.0, 57.8, 56.0, 55.8,55.0, 42.3, 41.4, 31.9, 29.7, 27.8, 26.9, 25.6, 24.0, 22.7, 20.5, 16.0,14.1, 13.6, 9.7.

ESI-MS m/z: 796.3 (M−H₂O+H)⁺.

(+)-HR-ESI-TOF-MS m/z: 796.3062 [M−H₂O+H]⁺ (Calcd. for C₄₂H₄₆N₅O₉S796.3011).

Example 13. Synthesis of allylN—[(R)-2-amino-3-(5-methoxy-1H-indol-3-yl)propyl)]carbamate (24-R)

To a solution of 17-R (2.35 g, 10.7 mmol) in CH₃CN (43 mL, 4 mL/mmol)was added di-tert-butyl dicarbonate (4.67 g, 21.4 mmol). The reactionmixture was stirred at 23° C. for 2.5 h, concentrated under vacuum.Flash chromatography (CH₂Cl₂:CH₃OH, from 99:1 to 85:15) afforded 20-R(1.7 g, 50%).

R_(f)=0.6 (CH₂Cl₂:CH₃OH, 9:1).

¹H NMR (400 MHz, CDCl₃): δ 8.05 (s, 1H), 7.25 (d, J=8.9 Hz, 1H), 7.09(d, J=2.4 Hz, 1H), 7.02 (d, J=2.4 Hz, 1H), 6.86 (dd, J=8.8, 2.4 Hz, 1H),4.83 (s, 1H), 3.98 (s, 1H), 3.87 (s, 3H), 3.69 (td, J=9.2, 7.5, 5.3 Hz,1H), 3.61 (dd, J=10.9, 5.6 Hz, 1H), 2.95 (d, J=6.8 Hz, 2H), 1.42 (s,9H).

To a solution of 20-R (1.7 g, 5.3 mmol) in CH₂Cl₂ (32 mL, 6 mL/mmol) wasadded phthalimide (1.72 g, 11.7 mmol), triphenylphosphine (3.06 g, 11.7mmol) and the mixture was cooled at 0° C. A solution of 40% of diethylazodicarboxylate in CH₂Cl₂ (4.0 mL, 13.2 mmol) was added for 15 min. Thereaction was stirred at 23° C. for 16 h, concentrated under vacuum. Theresidue obtained was purified by flash chromatography (Hexane:EtOAc,from 99:1 to 85:15) to afford 21-R (2.0 g, 84%).

R_(f)=0.45 (Hexane:EtOAc, 1:1).

¹H NMR (400 MHz, CDCl₃): δ 8.31 (s, 1H), 7.80 (dd, J=5.4, 3.0 Hz, 2H),7.67 (dd, J=5.4, 3.0 Hz, 2H), 7.30-7.12 (m, 2H), 7.08 (dd, J=15.2, 2.4Hz, 1H), 6.84 (dd, J=8.8, 2.4 Hz, 1H), 4.85 (d, J=9.2 Hz, 1H), 4.43 (q,J=5.3 Hz, 1H), 3.86 (s, 3H), 3.83-3.68 (m, 2H), 3.01 (d, J=5.4 Hz, 2H),1.22 (s, 9H).

To a solution of 21-R (2.0 g, 4.45 mmol) in ethanol (133 mL, 30 mL/mmol)was added hydrazine monohydrate (21.6 mL, 445 mmol). The reactionmixture was stirred at 80° C. in sealed tube for 2 h, concentrated undervacuum. Flash chromatography (CH₂Cl₂:CH₃OH, from 100:1 to 50:50) toafford 22-R (1.15 g, 81%).

R_(f)=0.1 (CH₂Cl₂:CH₃OH, 8:2).

¹H NMR (400 MHz, CDCl₃): δ 7.21 (d, J=8.8 Hz, 1H), 7.12 (s, 1H), 7.05(s, 1H), 6.75 (dd, J=8.8, 2.4 Hz, 1H), 3.95 (ddd, J=10.7, 8.7, 5.4 Hz,1H), 3.82 (s, 3H), 2.98-2.79 (m, 3H), 2.75 (dd, J=13.1, 9.4 Hz, 1H),1.37 (s, 9H).

To a solution of 22-R (1.1 g, 3.4 mmol) in CH₃CN (34 mL, 10 mL/mmol) andDMF (3.4 mL, 1 mL/mmol) was added N,N-diisopropylethylamine (0.5 mL, 2.7mmol) and allyl chloroformate (3.7 mL, 34 mmol). The reaction wasstirred at 23° C. for 19 h. The mixture was diluted with EtOAc and NH₄Clwas added and the mixture was extracted with EtOAc. The combined organiclayers were dried over anhydrous Na₂SO₄, filtered, and concentratedunder vacuum. The residue obtained was purified by flash chromatography(Hexane:EtOAc, from 100:1 to 1:100) to afford 23-R (0.95 g, 69%).

R_(f)=0.5 (Hexane:EtOAc, 1:1).

¹H NMR (400 MHz, CDCl₃): δ 8.55 (s, 1H), 7.20 (d, J=8.8 Hz, 1H), 7.05(s, 1H), 6.98-6.87 (m, 1H), 6.82 (dt, J=8.8, 1.8 Hz, 1H), 5.96-5.81 (m,1H), 5.37-5.22 (m, 2H), 5.22-5.14 (m, 1H), 5.02-4.97 (m, 1H), 4.60-4.47(m, 2H), 4.00 (s, 1H), 3.84 (s, 3H), 3.31 (s, 1H), 3.19 (s, 1H), 2.88(td, J=14.5, 13.3, 5.9 Hz, 2H), 1.40 (s, 9H).

To a solution of 23-R (0.94 g, 2.3 mmol) in CH₂Cl₂ (39 mL, 16.6 mL/mmol)was added trifluoroacetic acid (19 mL, 8.3 mL/mmol). The reactionmixture was stirred at 23° C. for 1.5 h, concentrated under vacuum toafford 24-R (0.72 g, 100%).

R_(f)=0.1 (CH₂Cl₂:CH₃OH, 9:1).

¹H NMR (400 MHz, CD₃OD): δ 7.27 (d, J=8.8, 1H), 7.18 (s, 1H), 7.04 (d,J=2.4 Hz, 1H), 6.80 (ddd, J=8.8, 2.4, 0.9 Hz, 1H), 5.95 (ddt, J=16.4,10.8, 5.5 Hz, 1H), 5.32 (d, J=17.1 Hz, 1H), 5.20 (d, J=10.5 Hz, 1H),4.60-4.53 (m, 2H), 3.83 (s, 3H), 3.59 (dt, J=11.4, 5.5 Hz, 1H),3.47-3.30 (m, 2H), 3.13-2.94 (m, 2H).

Example 14

To a solution of 1 (0.71 g, 1.14 mmol) in acetic acid (45 mL, 0.08 M)was added 24-R (0.54 mg, 1.8 mmol). The reaction mixture was stirred at23° C. for 7 h and then acetic acid was evaporated. An aqueous saturatedsolution of NaHCO₃ was added and the mixture was extracted with CH₂Cl₂.The combined organic layers were dried over anhydrous Na₂SO₄, filtered,and concentrated under vacuum. Flash chromatography (Hexane:EtOAc, 1:1)gives compound 25-R (670 mg, 65%).

R_(f)=0.4 (Hexane:EtOAc, 1:1).

¹H NMR (400 MHz, CDCl₃): δ 7.52 (s, 1H), 7.17 (d, J=8.8 Hz, 1H),6.83-6.73 (m, 2H), 6.61 (s, 1H), 6.23 (d, J=1.0 Hz, 1H), 6.02 (d, J=1.0Hz, 1H), 6.05-5.89 (m, 1H), 5.75 (s, 1H), 5.44-5.30 (m, 1H), 5.25 (d,J=10.4 Hz, 1H), 5.13-4.99 (m, 2H), 4.71-4.59 (m, 2H), 4.36 (s, 1H),4.30-4.07 (m, 3H), 3.80 (s, 3H), 3.79 (s, 3H), 3.61-3.53 (m, 1H);3.48-3.41 (m, 3H), 3.26 (dt, J=13.3, 3.8 Hz, 1H), 3.04-2.88 (m, 2H),2.52 (dd, J=14.9, 3.7 Hz, 1H), 2.46-2.35 (m, 2H), 2.31 (s, 3H), 2.29 (s,3H), 2.16 (s, 3H), 2.12-2.02 (m, 1H), 2.09 (s, 3H).

ESI-MS m/z: 907.3 (M+H)⁺.

To a solution of 25-R (745 mg, 0.82 mmol) in CH₂Cl₂ (15 mL, 18 mL/mmol)was added bis(triphenylphosphine)palladium(II) dichloride (92 mg, 0.1mmol) and acetic acid (0.47 mL, 8.2 mmol). Tributyltin hydride (1.33 mL,4.9 mmol) was added at 0° C., the reaction mixture was stirred at 0° C.for 0.75 h and concentrated under vacuum. Flash chromatography(Hexane:EtOAc, from 100:1 to 1:100 and EtOAc:CH₃OH, from 100:1 to 1:100)to afford 26-R (680 mg, >100%).

R_(f)=0.25 (CH₂Cl₂:CH₃OH, 1:1).

¹H NMR (400 MHz, CDCl₃): δ 7.57 (s, 1H), 7.16 (d, J=8.8 Hz, 1H),6.85-6.72 (m, 2H), 6.57 (s, 1H), 6.21 (d, J=1.4 Hz, 1H), 6.00 (d, J=1.3Hz, 1H), 5.05-4.97 (m, 1H), 4.63 (s, 1H), 4.35 (s, 1H), 4.31-4.09 (m,4H), 3.80 (s, 3H), 3.78 (s, 3H), 3.50-3.40 (m, 3H), 3.24 (dq, J=9.9, 5.3Hz, 1H), 2.95 (s, 1H), 2.91-2.75 (m, 2H), 2.62 (dd, J=14.8, 3.6 Hz, 1H),2.43-2.28 (m, 2H), 2.36 (s, 3H), 2.25 (s, 3H), 2.22-2.14 (m, 1H), 2.15(s, 3H), 2.08 (s, 3H).

ESI-MS m/z: 823.3 (M+H)⁺.

To a solution of 26-R (660 mg, 0.80 mmol) in CH₃CN:H₂O (1.39:1, 56 mL,0.015 M) was added AgNO₃ (2.70 g, 16.0 mmol). After 16.5 h at 23° C.,the reaction was quenched with a mixture 1:1 of saturated aqueoussolutions of NaCl and NaHCO₃, stirred for 15 min, diluted with CH₂Cl₂,stirred for 5 min, and extracted with CH₂Cl₂. The combined organiclayers were dried over anhydrous Na₂SO₄, filtered, and concentratedunder vacuum. The residue obtained was purified by flash chromatography(CH₂Cl₂:CH₃OH, from 99:1 to 85:15) to give 27-R (271 mg, 42%).

R_(f)=0.1 (CH₂Cl₂:CH₃OH, 9:1).

¹H NMR (400 MHz, CDCl₃): δ 7.46 (s, 1H), 7.16 (d, J=8.9 Hz, 1H), 6.83(s, 1H), 6.72 (d, J=8.9 Hz, 1H), 6.58 (s, 1H), 6.20 (d, J=1.8 Hz, 1H),5.99 (d, J=1.8 Hz, 1H), 5.76 (s, 1H), 5.15 (d, J=11.4 Hz, 1H), 4.86 (s,1H), 4.52 (m, 2H), 4.17 (d, J=5.3 Hz, 1H), 4.07 (d, J=11.4 Hz, 1H), 3.80(s, 3H), 3.78 (s, 3H), 3.55-3.43 (m, 2H), 3.32-3.20 (m, 2H), 3.01-2.82(m, 4H), 2.68-2.59 (m, 1H), 2.44-2.31 (m, 1H), 2.38 (s, 3H), 2.30-2.19(m, 1H), 2.26 (s, 3H), 2.15 (s, 3H), 2.07 (s, 3H).

¹³C NMR (101 MHz, CD₃OD): δ 171.7, 171.3, 153.8, 153.3, 148.0, 147.6,145.4, 145.4, 143.1, 141.3, 140.7, 131.6, 131.4, 131.2, 129.3, 126.8,121.6, 120.9, 118.3, 115.6, 112.2, 111.8, 101.8, 100.2, 81.7, 63.5,63.1, 61.7, 58.0, 57.8, 56.1, 55.8, 55.0, 42.2, 42.1, 41.4, 41.0, 25.1,23.8, 20.5, 16.0, 9.7.

ESI-MS m/z: 796.3 (M−H₂O+H)⁺.

(+)-HR-ESI-TOF-MS m/z: 796.3045 [M−H₂O+H]⁺ (Calcd. for C₄₂H₄₆N₅O₉S796.3011).

Example 15. Synthesis of Reference Compounds 28-S and 29-S

To a solution of 1 (450 mg, 0.72 mmol) in acetic acid (9 mL, 0.08 M) wasadded 16-S (675 mg, 2.88 mmol). The reaction mixture was stirred a 52°C. for 3 h and then acetic acid was evaporated. An aqueous saturatedsolution of NaHCO₃ was added and the mixture was extracted with CH₂Cl₂.The combined organic layers were dried over anhydrous Na₂SO₄, filtered,and concentrated under vacuum. Flash chromatography (CH₂Cl₂:CH₃OH, from99:1 to 80:20) gave compound 28-S (400 mg, 66%).

R_(f)=0.35 (CH₂Cl₂:CH₃OH, 10:1).

¹H NMR (400 MHz, CDCl₃): δ 7.65 (s, 1H), 7.15 (d, J=8.7 Hz, 1H),6.85-6.76 (m, 2H), 6.57 (s, 1H), 6.25 (d, J=1.4 Hz, 1H), 6.04 (d, J=1.3Hz, 1H), 5.16 (d, J=11.7 Hz, 1H), 4.62 (s, 1H), 4.44 (s, 1H), 4.35 (dd,J=11.7, 2.0 Hz, 1H), 4.29 (dd, J=5.2, 1.6 Hz, 1H), 4.22 (d, J=2.7 Hz,1H), 3.80 (s, 3H), 3.79 (s, 3H), 3.52-3.43 (m, 3H), 3.02-2.81 (m, 4H),2.41-2.31 (m, 2H), 2.36 (s, 3H), 2.29 (s, 3H), 2.15 (s, 3H), 2.05 (s,3H).

ESI-MS m/z: 838.6 (M+H)⁺.

To a solution of 28-S (400 mg, 0.48 mmol) in CH₃CN:H₂O (2:1, 33 mL,0.015 M) was added AgNO₃ (1.20 g, 7.16 mmol). After 16 h at 23° C., amixture 1:1 of saturated aqueous solutions of NaCl and NaHCO₃ was added,stirred for 15 min, diluted with CH₂Cl₂, stirred for 30 min, andextracted with CH₂Cl₂. The combined organic layers were dried overanhydrous Na₂SO₄, filtered, and concentrated under vacuum. The residueobtained was purified by flash chromatography (CH₂Cl₂:CH₃OH, from 99:1to 70:30) to afford 29-S (179 mg, 45%).

R_(f)=0.25 (CH₂Cl₂:CH₃OH, 9:1).

¹H NMR (500 MHz, CD₃OD): δ 7.17 (d, J=8.9 Hz, 1H), 6.83 (d, J=2.4 Hz,1H), 6.70 (dd, J=8.9, 2.4 Hz, 1H), 6.66 (s, 1H), 6.29 (d, J=1.3 Hz, 1H),6.10 (d, J=1.3 Hz, 1H), 5.32 (d, J=11.6 Hz, 1H), 4.65 (s, 1H), 4.57 (s,1H), 4.48 (s, 1H), 4.38 (dd, J=11.7, 2.1 Hz, 1H), 3.75 (s, 3H), 3.73 (s,3H), 3.41-3.35 (m, 1H), 3.16-2.91 (m, 5H), 2.71 (dd, J=15.3, 11.4 Hz,2H), 2.54 (s, 1H), 2.42-2.36 (m, 2H), 2.38 (s, 3H), 2.37 (s, 3H), 2.28(s, 3H), 1.99 (s, 3H).

¹³C NMR (126 MHz, CDCl₃): δ 171.3, 170.6, 155.2, 149.8, 147.5, 145.4,142.8, 142.4, 133.0, 131.8, 130.0, 128.0, 122.2, 121.8, 115.5, 113.9,113.3, 113.2, 111.4, 109.1, 103.8, 100.9, 91.6, 65.4, 61.9, 60.3, 59.4,57.1, 56.4, 56.2, 55.2, 53.4, 43.7, 40.8, 38.3, 30.7, 26.4, 24.7, 20.4,16.5, 9.6.

ESI-MS m/z: 811.3 (M−H₂O+H)⁺.

(+)-HR-ESI-TOF-MS m/z: 811.2682 [M−H₂O+H]⁺ (Calcd. for C₄₂H₄₃N₄O₁₁S811.2644).

Example 16. Synthesis of Reference Compounds 28-R and 29-R

To a solution of 1 (50 mg, 0.08 mmol) in acetic acid (1 mL, 0.08 M) wasadded 16-R (66 mg, 0.3 mmol). The reaction mixture was stirred at 50° C.for 6 h and then acetic acid was evaporated. An aqueous saturatedsolution of NaHCO₃ was added and the mixture was extracted with CH₂Cl₂.The combined organic layers were dried over anhydrous Na₂SO₄, filtered,and concentrated under vacuum. Flash chromatography (CH₂Cl₂:CH₃OH, from99:1 to 80:20) gave compound 28-R (50 mg, 75%).

R_(f)=0.20 (CH₂Cl₂:CH₃OH, 10:1).

¹H NMR (400 MHz, CDCl₃): 7.63 (s, 1H), 7.16 (d, J=8.8 Hz, 1H), 6.81 (d,J=2.4 Hz, 1H), 6.77 (dd, J=8.8, 2.3 Hz, 1H), 6.56 (s, 1H), 6.21 (d,J=1.2 Hz, 1H), 6.00 (d, J=1.2 Hz, 1H), 5.77 (s, 1H), 5.00 (d, J=11.8 Hz,1H), 4.63 (s, 1H), 4.35 (s, 1H), 4.27 (d, J=5.0 Hz, 1H), 4.22-4.04 (m,3H), 3.79 (s, 3H), 3.77 (s, 3H), 3.48-3.40 (m, 2H), 3.00 (dd, J=15.3,4.8 Hz, 1H), 2.92 (d, J=5.4 Hz, 2H), 2.71 (dd, J=15.3, 10.1 Hz, 1H),2.46 (d, J=14.9 Hz, 1H), 2.34 (s, 3H), 2.26 (s, 3H), 2.21 (d, J=15.0 Hz,1H), 2.15 (s, 3H), 2.07 (s, 3H).

ESI-MS m/z: 838.8 (M+H)⁺.

To a solution of 28-R (50 mg, 0.06 mmol) in CH₃CN:H₂O (2:1, 4.2 mL,0.015M) was added AgNO₃ (304 mg, 1.80 mmol). After 3 h at 23° C., amixture 1:1 of saturated aqueous solutions of NaCl and NaHCO₃ was added,stirred for 15 min, diluted with CH₂Cl₂, stirred for 30 min, andextracted with CH₂Cl₂. The combined organic layers were dried overanhydrous Na₂SO₄, filtered, and concentrated under vacuum. The residueobtained was purified by flash chromatography (CH₂Cl₂:CH₃OH from 99:1 to70:30) to afford 29-R (30 mg, 60%).

R_(f)=0.15 (CH₂Cl₂:CH₃OH, 9:1).

¹H NMR (400 MHz, CDCl₃): 7.68 (s, 1H), 7.14 (d, J=8.8 Hz, 1H), 6.80 (d,J=2.4 Hz, 1H), 6.76 (dd, J=8.8, 2.4 Hz, 1H), 6.57 (s, 1H), 6.17 (d,J=1.3 Hz, 1H), 5.95 (d, J=1.3 Hz, 1H), 5.75 (s, 1H), 5.12 (d, J=11.5 Hz,1H), 4.85 (s, 1H), 4.56-4.46 (m, 2H), 4.17 (s, 1H), 4.10 (dd, J=9.9, 4.9Hz, 1H), 4.05 (dd, J=11.4, 2.0 Hz, 1H), 3.78 (s, 3H), 3.76 (s, 3H), 3.51(s, 1H), 3.48-3.42 (m, 2H), 3.23 (s, 1H), 3.00 (dd, J=15.3, 4.9 Hz, 1H),2.90-2.77 (m, 2H), 2.71 (dd, J=15.2, 9.9 Hz, 1H), 2.48 (d, J=14.6 Hz,1H), 2.34 (s, 3H), 2.25 (s, 3H), 2.20 (d, J=14.6 Hz, 1H), 2.14 (s, 3H),2.05 (s, 3H).

¹³C NMR (101 MHz, CDCl₃): 175.6, 171.0, 168.7, 154.1, 147.3, 145.6,143.1, 141.3, 140.8, 131.1, 130.4, 126.5, 121.9, 121.5, 121.3, 115.5,112.9, 112.7, 112.0, 109.1, 101.9, 100.2, 81.5, 62.8, 61.7, 60.4, 57.9,57.8, 56.0, 55.8, 54.8, 53.4, 42.5, 41.2, 40.3, 29.7, 24.6, 23.8, 20.5,15.9, 9.8.

ESI-MS m/z: 811.6 (M−H₂O+H)⁺.

(+)-HR-ESI-TOF-MS m/z: 811.2687 [M−H₂O+H]⁺ (Calcd. for C₄₂H₄₃N₄O₁₁S811.2644).

Example 17

To a solution of compound 1 (2.0 g, 3.21 mmol) in acetonitrile (200 mL,0.01 M) was added 2-benzofuran-3-yl-ethylamine hydrochloride (30) (1.90g, 9.65 mmol, Sigma Aldrich) and cyanuric chloride (TCT) (200 mg, 10%).The reaction mixture was stirred at 85° C. for 24 h and then aqueoussaturated solution of NaHCO₃ was added and the mixture was extractedwith CH₂Cl₂. The combined organic layers were dried over anhydrousNa₂SO₄, filtered, and concentrated under vacuum. Flash chromatography(Hexane:EtOAc, from 9:1 to 1:9) gives compound 31 (1.95 g, 79%).

R_(f)=0.5 (Hexane:EtOAc, 1:1).

¹H NMR (400 MHz, CDCl₃): δ 7.38-7.36 (m, 2H), 7.19-7.10 (m, 2H), 6.64(s, 1H), 6.20 (d, J=1.5 Hz, 1H), 6.05 (d, J=1.5 Hz, 1H), 5.76 (s, 1H),5.05 (d, J=11.7 Hz, 1H), 4.54 (s, 1H), 4.33-4.24 (m, 2H), 4.23-4.16 (m,2H), 3.81 (s, 3H), 3.49-3.38 (m, 2H), 3.28-3.21 (m, 1H), 3.06-2.78 (m,5H), 2.57-2.50 (m, 2H), 2.37 (s, 3H), 2.27 (s, 3H), 2.21 (m, 3H), 2.08(s, 3H).

ESI-MS m/z: 765.3 (M+H)⁺.

To a solution of compound 31 (380 mg, 0.49 mmol) in CH₃CN:H₂O (1.39:1,25 mL, 0.015 M) was added AgNO₃ (1.30 g, 7.45 mmol). After 5 h at 23°C., a mixture 1:1 of saturated aqueous solutions of NaCl and NaHCO₃ wasadded, stirred for 15 min, diluted with CH₂Cl₂, stirred for 5 min, andextracted with CH₂Cl₂. The combined organic layers were dried overanhydrous Na₂SO₄, filtered, and concentrated under vacuum. The residueobtained was purified by flash chromatography (CH₂Cl₂:CH₃OH, from 99:1to 85:15) to afford compound 32 (175 mg, 47%).

R_(f)=0.40 (CH₂Cl₂:CH₃OH, 9:1).

¹H NMR (400 MHz, CDCl₃): δ 7.35 (ddd, J=10.7, 7.6, 1.1 Hz, 2H), 7.14(dtd, J=19.7, 7.3, 1.3 Hz, 2H), 6.65 (s, 1H), 6.16 (d, J=1.5 Hz, 1H),6.01 (d, J=1.5 Hz, 1H), 5.75 (s, 1H), 5.15 (dd, J=11.5, 1.2 Hz, 1H),4.80 (s, 1H), 4.48 (d, J=3.2 Hz, 1H), 4.44 (s, 1H), 4.20-4.06 (m, 2H),3.81 (s, 1H), 3.50 (d, J=18.8 Hz, 1H), 3.30 (ddd, J=12.6, 7.9, 5.1 Hz,1H), 3.22 (d, J=9.1 Hz, 1H), 2.99 (d, J=17.9 Hz, 1H), 2.84 (dd, J=19.2,12.0 Hz, 3H), 2.59-2.49 (m, 2H), 2.36 (s, 3H), 2.27 (s, 3H), 2.21-2.14(m, 1H), 2.18 (s, 3H), 2.06 (s, 3H).

¹³C NMR (101 MHz, CDCl₃): δ 171.2, 168.7, 154.4, 150.0, 147.9, 145.5,142.9, 140.9, 140.8, 131.3, 129.0, 127.7, 123.7, 122.2, 121.2, 120.8,118.9, 118.3, 115.5, 113.5, 111.7, 101.7, 82.1, 62.7, 61.7, 60.3, 57.8,57.4, 55.9, 55.0, 42.2, 41.3, 39.7, 38.2, 29.7, 23.7, 21.3, 20.6, 15.9,9.7.

ESI-MS m/z: 738.6 (M−H₂O+H)⁺.

(+)-HR-ESI-TOF-MS m/z: 756.2654 [M+H]⁺ (Calcd. for C₄₀H₄₂N₃O₁₀S756.2585).

Example 18

To a solution of 1 (500 mg, 0.80 mmol) in acetic acid (10 mL, 0.08 M)was added 2-(5-methoxybenzofuran-3-yl)-ethylamine hydrochloride (33)(Diverchim, ref: DW04590) (444 mg, 1.60 mmol). The reaction mixture wasstirred at 50° C. for 6 days and then acetic acid was evaporated. Anaqueous saturated solution of NaHCO₃ was added and the mixture wasextracted with CH₂Cl₂. The combined organic layers were dried overanhydrous Na₂SO₄, filtered, and concentrated under vacuum. Flashchromatography (Hexane:EtOAc, 1:1) affords 34 (270 mg, 43%).

R_(f)=0.3 (Hexane:EtOAc, 1:1).

¹H NMR (400 MHz, CDCl₃): δ 7.25 (d, J=9.1 Hz, 1H), 6.80-6.73 (m, 2H),6.63 (s, 1H), 6.18 (d, J=1.4 Hz, 1H), 6.03 (d, J=1.4 Hz, 1H), 5.78 (s,1H), 5.03 (dd, J=11.5, 1.3 Hz, 1H), 4.52 (s, 1H), 4.29 (s, 1H), 4.26(dd, J=4.7, 1.5 Hz, 1H), 4.23-4.16 (m, 2H), 3.80 (s, 3H), 3.78 (s, 3H),3.46-3.43 (m, 1H), 3.43-3.37 (m, 1H), 3.24 (s, 1H), 3.03 (d, J=18.0 Hz,1H), 2.91 (dd, J=17.9, 9.2 Hz, 1H), 2.87-2.72 (m, 2H), 2.53-2.47 (m,2H), 2.36 (s, 3H), 2.27 (s, 3H), 2.20 (s, 3H), 2.06 (s, 3H).

ESI-MS m/z: 795.8 (M+H)⁺.

To a solution of 34 (345 mg, 0.43 mmol) in CH₃CN:H₂O (1.39:1, 30 mL,0.015 M) was added AgNO₃ (2.20 g, 13.0 mmol). After 3 h at 23° C., amixture 1:1 of saturated aqueous solutions of NaCl and NaHCO₃ was added,stirred for 15 min, diluted with CH₂Cl₂, stirred for 5 min, andextracted with CH₂Cl₂. The combined organic layers were dried overanhydrous Na₂SO₄, filtered, and concentrated under vacuum. The residueobtained was purified by flash chromatography (CH₂Cl₂:CH₃OH, from 99:1to 85:15) to obtain 35 (175 mg, 51%).

R_(f)=0.35 (CH₂Cl₂:CH₃OH, 9:1).

¹H NMR (500 MHz, CD₃OD): δ 7.27 (d, J=9.0 Hz, 1H), 6.90 (d, J=2.6 Hz,1H), 6.80 (dd, J=9.0, 2.6 Hz, 1H), 6.57 (s, 1H), 6.23 (d, J=1.2 Hz, 1H),6.05 (d, J=1.2 Hz, 1H), 5.23 (d, J=11.5 Hz, 1H), 4.27-4.08 (m, 4H), 3.77(s, 3H), 3.75 (s, 3H), 3.63 (d, J=14.1 Hz, 2H), 3.40-3.34 (m, 2H),2.93-2.87 (m, 5H), 2.80 (d, J=15.5 Hz, 1H), 2.57-2.54 (m, 2H), 2.34 (s,3H), 2.30 (s, 3H), 2.14 (s, 3H), 2.05 (s, 3H).

¹³C NMR (126 MHz, CD₃OD): δ 171.9, 170.6, 157.5, 147.0, 145.0, 142.3,141.0, 132.2, 131.1, 129.1, 122.2, 120.9, 120.2, 116.3, 115.1, 114.0,112.7, 111.4, 103.5, 102.7, 92.9, 62.0, 60.3, 59.8, 59.4, 56.5, 56.2,56.0, 54.0, 43.8, 41.2, 40.7, 30.8, 30.3, 28.7, 24.5, 21.6, 20.6, 16.2,9.6.

ESI-MS m/z: 768.6 (M−H₂O+H)⁺.

(+)-HR-ESI-TOF-MS m/z: 768.2630 [M−H₂O+H]⁺ (Calcd. for C₄₁H₄₂N₃O₁₀S768.2585).

Example 19

To a solution of LiAlH₄ (148 mL, 1.0 M in THF, 148 mmol) at −40° C. wasadded carefully H₂SO₄ (7.14 mL, 72.9 mmol) and a suspension of(S)-2-amino-3-(benzofuran-3-yl)propanoic acid (36-S) (prepared asdescribed in Tetrahedron Asymmetry 2008, 19, 500-511) (5.54 g, 26.9mmol) in THF (85 mL, 0.003 M). The reaction mixture was left evolutionat 23° C., heated at 80° C. for 3 h and 18 h at 23° C. Cool at −21° C.the reaction mixture was quenched carefully with NaOH 2N until basic pH.EtOAc was added and the mixture filtered through Celite® and washed withCH₃OH. The crude was concentrated under vacuum to afford compound 37-S(3.93 g, >100%).

R_(f)=0.1 (CH₂Cl₂:CH₃OH, 4:1).

¹H NMR (400 MHz, CD₃OD): δ 7.67-7.62 (m, 1H), 7.61 (s, 1H), 7.51-7.41(m, 1H), 7.34-7.18 (m, 2H), 3.69-3.48 (m, 1H), 3.44 (dd, J=10.8, 6.6 Hz,1H), 3.18 (dtd, J=7.4, 6.4, 4.6 Hz, 1H), 2.88 (ddd, J=14.4, 6.1, 1.0 Hz,1H), 2.68 (ddd, J=14.4, 7.5, 0.9 Hz, 1H).

Example 20

To a solution of LiAlH₄ (118 mL, 1.0 M in THF, 118 mmol) at −40° C. wasadded carefully H₂SO₄ (3.1 mL, 57.8 mmol) and a suspension of(R)-2-amino-3-(benzofuran-3-yl)propanoic acid (36-R) (prepared asdescribed in Tetrahedron Asymmetry 2008, 19, 500-511) (4.4 g, 21.4 mmol)in THF (67.4 mL, 0.003 M). The reaction mixture was left evolution at23° C., heated at 80° C. for 3 h and 18 h at 23° C. Cool at −21° C. thereaction mixture was quenched carefully with NaOH 2N until basic pH.EtOAc was added and the mixture filtered through Celite® and washed withCH₃OH. The crude was concentrated under vacuum. Flash chromatography(CH₂Cl₂:CH₃OH, from 99:1 to 85:15, Silice amine) to afford compound 37-R(2.77 g, 68%).

R_(f)=0.1 (CH₂Cl₂:CH₃OH, 4:1).

¹H NMR (400 MHz, CD₃OD): δ 7.63-7.52 (m, 1H), 7.56 (s, 1H), 7.46-7.33(m, 1H), 7.21 (dtd, J=19.9, 7.3, 1.3 Hz, 2H), 3.57 (dd, J=10.7, 4.6 Hz,1H), 3.42 (dd, J=10.8, 6.6 Hz, 1H), 3.15 (dtd, J=7.6, 6.3, 4.6 Hz, 1H),2.84 (ddd, J=14.4, 6.0, 1.0 Hz, 1H), 2.64 (ddd, J=14.4, 7.5, 0.9 Hz,1H).

Example 21

To a solution of compound 1 (850 mg, 1.36 mmol) in CH₃CN (136 mL, 0.01M) was added (S)-2-amino-3-(benzofuran-3-yl)propan-1-ol (37-S) (1.30 g,6.83 mmol and cyanuric chloride (TCT) (170 mg, 20%). The reactionmixture was stirred at 85° C. for 24 h and then aqueous saturatedsolution of NaHCO₃ was added and the mixture was extracted with CH₂Cl₂.The combined organic layers were dried over anhydrous Na₂SO₄, filtered,and concentrated under vacuum. Flash chromatography (Hexane:EtOAc, from9:1 to 1:9) gives compound 38-S (750 mg, 69%).

R_(f)=0.25 (Hexane:EtOAc, 1:1).

¹H NMR (400 MHz, CDCl₃): δ 7.39-7.33 (m, 1H), 7.33-7.29 (m, 1H), 7.20(ddd, J=8.3, 7.2, 1.4 Hz, 1H), 7.14 (td, J=7.4, 1.0 Hz, 1H), 6.61 (s,1H), 6.21 (d, J=1.4 Hz, 1H), 6.06 (d, J=1.4 Hz, 1H), 5.74 (s, 1H), 5.08(d, J=11.2 Hz, 1H), 4.58 (s, 1H), 4.37 (s, 1H), 4.32-4.23 (m, 2H), 4.19(d, J=2.7 Hz, 1H), 3.81 (s, 3H), 3.52-3.41 (m, 3H), 3.36-3.29 (m, 1H),3.13 (d, J=9.8 Hz, 1H), 3.00-2.81 (m, 3H), 2.57 (dd, J=15.7, 4.9 Hz,1H), 2.50 (d, J=15.2 Hz, 1H), 2.37 (s, 3H), 2.31-2.25 (m, 1H), 2.29 (s,3H), 2.16 (s, 3H), 2.10 (d, J=7.2 Hz, 1H), 2.05 (s, 3H).

ESI-MS m/z: 795.2 (M)⁺.

To a solution of compound 38-S (890 mg, 1.12 mmol) in CH₃CN:H₂O (1.39:1,75 mL, 0.015 M) was added AgNO₃ (4.70 g, 28.0 mmol). After 18 h at 23°C., a mixture 1:1 of saturated aqueous solutions of NaCl and NaHCO₃ wasadded, stirred for 15 min, diluted with CH₂Cl₂, stirred for 5 min, andextracted with CH₂Cl₂. The combined organic layers were dried overanhydrous Na₂SO₄, filtered, and concentrated under vacuum. The residueobtained was purified by flash chromatography (CH₂Cl₂:CH₃OH, from 99:1to 85:15) to afford compound 39-S (500 mg, 57%).

R_(f)=0.30 (CH₂Cl₂:CH₃OH, 9:1). ¹H NMR (400 MHz, CDCl₃): δ 7.38-7.33 (m,1H), 7.33-7.28 (m, 1H), 7.23-7.16 (m, 1H), 7.16-7.09 (m, 1H), 6.62 (s,1H), 6.18 (d, J=1.4 Hz, 1H), 6.03 (d, J=1.4 Hz, 1H), 5.71 (s, 1H), 5.19(d, J=11.2 Hz, 1H), 4.85 (s, 1H), 4.49 (s, 2H), 4.24-4.10 (m, 3H), 3.81(s, 3H), 3.54 (d, J=4.9 Hz, 1H), 3.49 (d, J=2.3 Hz, 3H), 3.33 (t, J=10.1Hz, 2H), 3.22 (s, 1H), 2.98 (s, 1H), 2.84 (d, J=7.6 Hz, 2H), 2.62-2.53(m, 2H), 2.37 (s, 3H), 2.30-2.24 (m, 1H), 2.28 (s, 3H), 2.14 (s, 3H),2.04 (s, 3H).

¹³C NMR (126 MHz, CDCl₃): δ 172.0, 170.7, 156.1, 150.6, 149.9, 147.1,145.0, 142.4, 142.2, 132.0, 131.4, 128.7, 125.5, 123.8, 122.6, 121.6,120.1, 116.5, 114.4, 112.3, 103.5, 92.6, 66.0, 65.1, 62.2, 60.4, 59.7,56.6, 56.1, 54.8, 54.1, 51.6, 44.0, 41.3, 38.3, 30.8, 24.8, 20.6, 16.3,9.6.

ESI-MS m/z: 768.2 (M−H₂O+H)⁺.

(+)-HR-ESI-TOF-MS m/z: 768.2652 [M−H₂O+H]⁺ (Calcd. for C₄₁H₄₂N₃O₁₀S768.2585)

Example 22

To a solution of compound 1 (100 mg, 0.16 mmol) in CH₃CN (16 mL, 0.01 M)was added (R)-2-amino-3-(benzofuran-3-yl)propan-1-ol (37-R) (307 mg, 1.6mmol) and cyanuric chloride (TCT) (40 mg, 40%). The reaction mixture wasstirred at 85° C. for 44 h and then aqueous saturated solution of NaHCO₃was added and the mixture was extracted with CH₂Cl₂. The combinedorganic layers were dried over anhydrous Na₂SO₄, filtered, andconcentrated under vacuum. Flash chromatography (Hexane:EtOAc, from 9:1to 1:9) gives compound 38-R (95 mg, 75%).

R_(f)=0.3 (Hexane:EtOAc, 1:1).

¹H NMR (400 MHz, CDCl₃): δ 7.42-7.27 (m, 2H), 7.28-7.09 (m, 2H), 6.58(s, 1H), 6.20 (d, J=1.4 Hz, 1H), 6.05 (d, J=1.4 Hz, 1H), 5.79 (s, 1H),5.00 (d, J=11.4 Hz, 1H), 4.59 (s, 1H), 4.34 (s, 1H), 4.31-4.16 (m, 4H),3.80 (s, 3H), 3.79-3.76 (m, 1H), 3.63 (s, 1H), 3.54-3.40 (m, 4H),2.99-2.87 (m, 2H), 2.68 (d, J=15.0 Hz, 1H), 2.56-2.47 (m, 1H), 2.38 (s,3H), 2.27 (s, 3H), 2.17 (s, 3H), 2.07 (s, 3H).

ESI-MS m/z: 795.2 (M+H)⁺.

To a solution of compound 38-R (95 mg, 0.11 mmol) in CH₃CN:H₂O (1.39:1,11 mL, 0.015 M) was added AgNO₃ (601 mg, 3.58 mmol). After 18 h at 23°C., a mixture 1:1 of saturated aqueous solutions of NaCl and NaHCO₃ wasadded, stirred for 15 min, diluted with CH₂Cl₂, stirred for 5 min, andextracted with CH₂Cl₂. The combined organic layers were dried overanhydrous Na₂SO₄, filtered, and concentrated under vacuum. The residueobtained was purified by flash chromatography (CH₂Cl₂:CH₃OH, from 99:1to 85:15) to afford compound 39-R (66 mg, 70%).

R_(f)=0.3 (CH₂Cl₂:CH₃OH, 9:1).

¹³C NMR (400 MHz, CDCl₃): δ 7.39-7.31 (m, 2H), 7.23-7.07 (m, 2H), 6.59(s, 1H), 6.17 (d, J=1.4 Hz, 1H), 6.01 (d, J=1.4 Hz, 1H), 5.75 (s, 1H),5.12 (dd, J=11.3, 1.2 Hz, 1H), 4.84 (s, 1H), 4.56-4.43 (m, 2H),4.19-4.07 (m, 3H), 3.79 (s, 3H), 3.83-3.74 (m, 1H), 3.66-3.51 (m, 3H),3.24 (s, 1H), 2.99-2.79 (m, 2H), 2.75-2.64 (m, 1H), 2.59-2.43 (m, 2H),2.38 (s, 3H), 2.27 (s, 3H), 2.16 (s, 3H), 2.07 (s, 3H).

¹³C NMR (101 MHz, CD₃OD): δ 170.5, 169.1, 154.9, 148.9, 148.5, 145.7,143.6, 141.1, 140.8, 130.6, 129.9, 127.1, 124.1, 122.4, 122.4, 121.2,120.3, 118.7, 118.2, 115.1, 113.6, 110.9, 102.1, 91.1, 65.0, 63.3, 60.2,59.0, 58.4, 55.4, 54.5, 52.7, 52.3, 42.5, 38.7, 29.4, 23.5, 23.2, 19.1,14.8, 8.3.

ESI-MS m/z: 768.2 (M−H₂O+H)⁺.

(+)-HR-ESI-TOF-MS m/z: 767.2628 [M−H₂O+H]⁺ (Calcd. for C₄₁H₄₂N₃O₁₀S768.2585).

Example 23. Synthesis ofallyl-N—[(S)-2-amino-3-(benzofuran-3-yl)propyl]carbamate (44-S)

To a solution of compound 37-S (1.0 g, 5.22 mmol) in CH₃CN (21 mL, 4mL/mmol) was added di-tert-butyl dicarbonate (2.28 g, 10.4 mmol). Thereaction mixture was stirred at 23° C. for 2 h, concentrated undervacuum. Flash chromatography (CH₂Cl₂:CH₃OH, from 99:1 to 85:15) toafford compound 40-5 (0.5 g, 33%).

R_(f)=0.7 (CH₂Cl₂:CH₃OH, 9:1).

¹H NMR (400 MHz, CDCl₃): δ 7.64 (d, J=7.6 Hz, 1H), 7.49 (s, 1H), 7.46(d, J=7.6 Hz, 1H), 7.36-7.19 (m, 2H), 4.94 (s, 1H), 3.98 (s, 1H),3.71-3.56 (m, 2H), 2.93 (d, J=6.9 Hz, 2H), 1.41 (s, 9H).

To a solution of compound 40-S (0.5 g, 1.71 mmol) in CH₂Cl₂ (11 mL, 6mL/mmol) was added phthalimide (0.55 g, 3.77 mmol), Triphenylphosphine(0.99 g, 3.77 mmol) and the mixture was cooled at 0° C. A solution of40% of Diethyl azodicarboxylate in CH₂Cl₂ (1.26 mL, 4.29 mmol) was addedfor 15 min. The reaction was stirred at 23° C. for 18 h, concentratedunder vacuum. The residue obtained was purified by flash chromatography(Hexane:EtOAc, from 99:1 to 40:60) to afford compound 41-S (0.68 g,94%).

R_(f)=0.8 (CH₂Cl₂:CH₃OH, 9:1).

¹H NMR (400 MHz, CDCl₃): δ 7.89-7.79 (m, 2H), 7.83-7.62 (m, 2H),7.65-7.55 (m, 2H), 7.49-7.42 (m, 1H), 7.33-7.20 (m, 2H), 4.83 (d, J=9.0Hz, 1H), 4.39 (ddt, J=12.1, 6.3, 2.9 Hz, 1H), 3.88-3.70 (m, 2H), 2.96(d, J=6.4 Hz, 2H), 1.24 (s, 9H).

To a solution of compound 41-5 (345 mg, 0.82 mmol) in ethanol (25 mL, 30mL/mmol) was added hydrazine monohydrate (3.6 mL, 73.8 mmol). Thereaction mixture was stirred at 80° C. in sealed tube for 2 h,concentrated under vacuum. Flash chromatography (CH₂Cl₂:CH₃OH, from100:1 to 50:50) to afford compound 42-S (233 mg, 98%).

R_(f)=0.1 (CH₂Cl₂:CH₃OH, 8:2).

¹H NMR (400 MHz, CDCl₃): δ 7.62 (d, J=7.5 Hz, 1H), 7.49-7.42 (m, 2H),7.33-7.18 (m, 2H), 4.85 (d, J=8.8 Hz, 1H), 3.91 (s, 1H), 2.91-2.76 (m,3H), 2.67 (dd, J=13.1, 6.8 Hz, 1H), 1.25 (s, 9H).

To a solution of compound 42-S (280 mg, 0.96 mmol) in CH₃CN (10 mL, 10mL/mmol) and DMF (16 mL, 1 mL/mmol) was added N,N-diisopropylethylamine(0.14 mL, 0.77 mmol) and allyl chloroformate (1.02 mL, 9.64 mmol). Thereaction was stirred at 23° C. for 2 h. The mixture was diluted withEtOAc and NH₄Cl was added and the mixture was extracted with EtOAc. Thecombined organic layers were dried over anhydrous Na₂SO₄, filtered, andconcentrated under vacuum. The residue obtained was purified by flashchromatography (Hexane:EtOAc, from 100:1 to 1:100) to afford compound43-S (445 mg, >100%).

R_(f)=0.5 (Hexane:EtOAc, 1:1).

¹H NMR (400 MHz, CDCl₃): δ 7.60 (d, J=7.6 Hz, 1H), 7.52-7.43 (m, 2H),7.34-7.20 (m, 2H), 5.90 (ddt, J=16.4, 10.8, 5.6 Hz, 1H), 5.32-5.17 (m,2H), 4.93-4.86 (m, 1H), 4.56 (d, J=5.6 Hz, 2H), 4.08-3.98 (m, 1H),3.40-3.21 (m, 2H), 2.88 (m, 2H), 1.25 (s, 9H).

To a solution of compound 43-S (160 mg, 0.43 mmol) in CH₂Cl₂ (8 mL, 16.6mL/mmol) was added trifluoroacetic acid (4 mL, 8.3 mL/mmol). Thereaction mixture was stirred at 23° C. for 1.5 h, concentrated undervacuum. Flash chromatography (CH₂Cl₂:CH₃OH, from 100:1 to 50:50) toafford compound 44-S (175 mg, >100%).

R_(f)=0.2 (CH₂Cl₂:CH₃OH, 9:1).

¹H NMR (400 MHz, CD₃OD): δ 7.72 (s, 1H), 7.64 (dt, J=8.4, 0.9 Hz, 1H),7.49 (dt, J=8.4, 0.9 Hz, 1H), 7.37-7.22 (m, 2H), 5.94 (ddt, J=16.3,10.7, 5.5 Hz, 1H), 5.32 (dq, J=17.3, 1.7 Hz, 1H), 5.19 (dq, J=10.6, 1.5Hz, 1H), 4.56 (dt, J=5.7, 1.5 Hz, 2H), 3.56 (qd, J=7.0, 4.4 Hz, 1H),3.46-3.32 (m, 1H), 3.32-3.24 (m, 1H), 3.03 (dd, J=14.8, 6.9 Hz, 1H),2.91 (ddd, J=14.8, 7.1, 0.9 Hz, 1H).

Example 24. Synthesis ofallyl-N—[(R)-2-amino-3-(benzofuran-3-yl)propyl]carbamate (44-R)

To a solution of compound 37-R (2.75 g, 14.4 mmol) in CH₃CN (58 mL, 4mL/mmol) was added di-tert-butyl dicarbonate (6.27 g, 28.76 mmol). Thereaction mixture was stirred at 23° C. for 2.5 h, concentrated undervacuum. Flash chromatography (CH₂Cl₂:CH₃OH, from 99:1 to 85:15) toafford compound 40-R (3.7 g, 88%).

R_(f)=0.6 (CH₂Cl₂:CH₃OH, 9:1).

¹H NMR (400 MHz, CDCl₃): δ 7.64 (d, J=7.6 Hz, 1H), 7.52-7.43 (m, 2H),7.35-7.20 (m, 2H), 4.85 (d, J=8.2 Hz, 1H), 4.00 (bs, 1H), 3.69 (dd,J=11.0, 4.0 Hz, 1H), 3.62 (dd, J=10.9, 5.1 Hz, 1H), 2.94 (d, J=6.9 Hz,2H), 1.42 (s, 9H).

To a solution of compound 40-R (3.7 g, 12.7 mmol) in CH₂Cl₂ (76 mL, 6mL/mmol) was added phthalimide (4.1 g, 28 mmol), triphenylphosphine (7.3g, 28 mmol) and the mixture was cooled at 0° C. A solution of 40% ofdiethyl azodicarboxylate in CH₂Cl₂ (9.4 mL, 31.7 mmol) was added for 15min. The reaction was stirred at 23° C. for 16 h, concentrated undervacuum. The residue obtained was purified by flash chromatography(CH₂Cl₂:CH₃OH, from 99:1 to 85:15) to afford compound 41-R (4.05 g,76%).

R_(f)=0.8 (CH₂Cl₂:CH₃OH, 9:1).

¹H NMR (400 MHz, CDCl₃): δ 7.67-7.68 (m, 4H), 7.61 (d, J=7.5 Hz, 1H),7.58 (s, 1H), 7.46 (d, J=7.5 Hz, 1H), 7.27 (dtd, J=17.2, 7.3, 1.4 Hz,2H), 4.84 (d, J=9.0 Hz, 1H), 4.46-4.30 (m, 1H), 3.89-3.66 (m, 2H), 2.97(d, J=6.4 Hz, 2H), 1.24 (s, 9H).

To a solution of compound 41-R (4.0 g, 9.5 mmol) in ethanol (285 mL, 30mL/mmol) was added hydrazine monohydrate (41.5 mL, 856 mmol). Thereaction mixture was stirred at 80° C. in sealed tube for 2 h,concentrated under vacuum. Flash chromatography (CH₂Cl₂:CH₃OH, from100:1 to 50:50) to afford compound 42-R (2.2 g, 80%).

R_(f)=0.1 (CH₂Cl₂:CH₃OH, 8:2).

¹H NMR (400 MHz, CDCl₃): δ 7.60 (d, J=7.5 Hz, 1H), 7.45 (s, 1H), 7.44(d, J=7.1 Hz, 1H), 7.25 (dtd, J=18.8, 7.3, 1.3 Hz, 2H), 4.94 (d, J=8.8Hz, 1H), 3.98-3.78 (m, 1H), 2.90-2.77 (m, 2H), 2.65 (dd, J=13.1, 7.0 Hz,1H), 1.40 (s, 9H).

To a solution of compound 42-R (2.2 g, 7.6 mmol) in CH₃CN (76 mL, 10mL/mmol) and DMF (7.6 mL, 1 mL/mmol) was added N,N-diisopropylethylamine(1.1 mL, 6.08 mmol) and allyl chloroformate (8.05 mL, 76 mmol). Thereaction was stirred at 23° C. for 7 h. The mixture was diluted withEtOAc and NH₄Cl was added and the mixture was extracted with EtOAc. Thecombined organic layers were dried over anhydrous Na₂SO₄, filtered, andconcentrated under vacuum. The residue obtained was purified by flashchromatography (Hexane:EtOAc, from 100:1 to 1:100) to afford compound43-R (2.3 g, 81%).

R_(f)=0.7 (Hexane:EtOAc, 1:1).

¹H NMR (400 MHz, CDCl₃): δ 7.60 (d, J=7.5 Hz, 1H), 7.52-7.43 (m, 2H),7.34-7.20 (m, 2H), 5.90 (ddt, J=17.3, 10.8, 5.6 Hz, 1H), 5.29 (d,J=17.2, 1H), 5.20 (d, J=10.4, 1H), 5.10 (t, J=6.2 Hz, 1H), 4.86 (d,J=8.4 Hz, 1H), 4.56 (d, J=5.4, 2H), 4.08-3.97 (m, 1H), 3.36 (dt, J=10.7,4.7 Hz, 1H), 3.30-3.23 (m, 1H), 2.87 (td, J=14.8, 6.5 Hz, 2H), 1.41 (s,9H).

To a solution of compound 43-R (1.32 g, 3.52 mmol) in CH₂Cl₂ (60 mL,16.6 mL/mmol) was added Trifluoroacetic acid (30 mL, 8.3 mL/mmol). Thereaction mixture was stirred at 23° C. for 1.5 h, concentrated undervacuum. Flash chromatography (CH₂Cl₂:CH₃OH, from 100:1 to 50:50) toafford compound 44-R (0.90 g, 94%).

R_(f)=0.2 (CH₂Cl₂:CH₃OH, 9:1).

¹H NMR (400 MHz, CDCl₃): δ 7.75 (s, 1H), 7.69-7.61 (m, 1H), 7.54-7.46(m, 1H), 7.39-7.24 (m, 2H), 5.95 (ddt, J=16.3, 10.8, 5.5 Hz, 1H), 5.32(dd, J=17.3, 1.8 Hz, 1H), 5.24-5.16 (m, 1H), 4.57 (dt, J=5.7, 1.5 Hz,2H), 3.68 (qd, J=7.1, 4.2 Hz, 1H), 3.48 (dd, J=14.8, 4.2 Hz, 1H),3.42-3.30 (m, 1H), 3.14-2.95 (m, 2H).

Example 25

To a solution of compound 1 (750 mg, 1.2 mmol) in CH₃CN (120 mL, 0.01 M)was added compound 44-S (1370 mg, 6 mmol) and cyanuric chloride (TCT)(184 mg, 20%). The reaction mixture was stirred at 85° C. for 23 h andthen aqueous saturated solution of NaHCO₃ was added and the mixture wasextracted with CH₂Cl₂. The combined organic layers were dried overanhydrous Na₂SO₄, filtered, and concentrated under vacuum. Flashchromatography (Hexane:EtOAc, from 9:1 to 1:9) gives compound 45-S (755mg, 72%).

R_(f)=0.36 (Hexane:EtOAc, 1:1).

¹H NMR (400 MHz, CDCl₃): δ 7.38-7.28 (m, 2H), 7.23-7.08 (m, 2H), 6.67(s, 1H), 6.19 (d, J=1.4 Hz, 1H), 6.09-5.95 (m, 1H), 6.04 (d, J=1.4 Hz,1H), 5.92 (s, 1H), 5.80 (s, 1H), 5.44-5.34 (m, 1H), 5.26 (dq, J=10.4,1.3 Hz, 1H), 5.08 (dd, J=11.4, 1.1 Hz, 1H), 4.70-4.63 (m, 2H), 4.56 (s,1H), 4.34 (s, 1H), 4.31-4.18 (m, 3H), 3.80 (s, 3H), 3.50-3.39 (m, 2H),3.24-3.15 (m, 1H), 3.00 (dt, J=12.2, 6.0 Hz, 2H), 2.95 (d, J=5.2 Hz,2H), 2.60 (dd, J=15.4, 4.5 Hz, 2H), 2.44 (dd, J=15.6, 5.2 Hz, 1H), 2.29(s, 3H), 2.27 (s, 3H), 2.25-2.20 (m, 1H), 2.18 (s, 3H), 2.12 (s, 1H),2.04 (s, 3H).

ESI-MS m/z: 878.2 (M+H)⁺.

To a solution of compound 45-S (750 mg, 0.85 mmol) in CH₂Cl₂ (15.3 mL,18 mL/mmol) was added bis(triphenylphosphine)palladium(II) dichloride(96 mg, 0.14 mmol) and acetic acid (0.5 mL, 8.5 mmol). Tributyltinhydride (1.4 mL, 5.1 mmol) was added at 0° C., and the reaction mixturewas stirred at 0° C. for 30 minutes, and was concentrated under vacuum.Flash chromatography (Hexane:EtOAc, from 100:1 to 1:100 andCH₂Cl₂:CH₃OH, from 100:1 to 1:100) to afford compound 46-S (430 mg,64%).

R_(f)=0.3 (CH₂Cl₂:CH₃OH, 1:1).

¹H NMR (400 MHz, CDCl₃): δ 7.37-7.29 (m, 2H), 7.22-7.11 (m, 2H), 6.57(s, 1H), 6.21 (d, J=1.5 Hz, 1H), 6.06 (d, J=1.5 Hz, 1H), 5.07 (d, J=11.5Hz, 1H), 4.57 (s, 1H), 4.37 (s, 1H), 4.29-4.23 (m, 2H), 4.14 (s, 1H),3.79 (s, 3H), 3.50-3.47 (m, 2H), 3.38 (d, J=8.7 Hz, 1H), 2.95-2.71 (m,4H), 2.68-2.52 (m, 2H), 2.51-2.38 (m, 1H), 2.35 (s, 3H), 2.33-2.26 (m,1H), 2.29 (s, 3H), 2.17-2.08 (m, 1H), 2.10 (s, 3H), 2.04 (s, 3H).

ESI-MS m/z: 794.3 (M+H)⁺.

To a solution of compound 46-S (550 mg, 0.7 mmol) in CH₃CN:H₂O (1.39:1,49 mL, 0.015 M) was added AgNO₃ (2.4 g, 14 mmol). After 16 h at 23° C.,the reaction was quenched with a mixture 1:1 of saturated aqueoussolutions of NaCl and NaHCO₃, stirred for 15 min, diluted with CH₂Cl₂,stirred for 5 min, and extracted with CH₂Cl₂. The combined organiclayers were dried over anhydrous Na₂SO₄, filtered, and concentratedunder vacuum. The residue obtained was purified by flash chromatography(CH₂Cl₂:CH₃OH, from 99:1 to 85:15) to give compound 47-5 (53 mg, 10%).

R_(f)=0.1 (CH₂Cl₂:CH₃OH, 9:1).

¹H NMR (500 MHz, CDCl₃): δ 7.36 (d, 7.9 Hz, 1H), 7.33 (d, 7.4 Hz, 1H),7.23 (t, J=7.4 Hz, 1H), 7.16 (t, J=7.4 Hz, 1H), 6.77 (s, 1H), 6.20 (s,1H), 6.04 (s, 1H), 5.92 (s, 1H), 5.20 (d, J=11.1 Hz, 1H), 4.90 (s, 1H),4.50 (s, 1H), 4.46-4.39 (m, 1H), 4.25 (d, J=11.1 Hz, 1H), 4.20 (s, 1H),3.84 (s, 3H), 3.81 (d, J=4.2 Hz, 1H), 3.58 (s, 1H), 3.40-3.14 (m, 3H),2.90 (t, J=13.0 Hz, 1H), 2.76 (m, 3H), 2.50 (s, 3H), 2.46-2.37 (m, 1H),2.32-2.26 (m, 2H), 2.30 (s, 3H), 2.15 (s, 3H), 2.04 (s, 3H).

¹³C NMR (126 MHz, CD₃OD): δ 170.5, 169.2, 154.6, 149.1, 148.7, 145.7,143.5, 141.0, 140.9, 131.2, 129.6, 126.9, 124.4, 122.5, 121.4, 119.7,118.7, 115.0, 112.7, 111.0, 110.7, 102.1, 91.2, 63.5, 61.2, 59.2, 58.5,55.3, 54.7, 53.4, 52.7, 43.3, 42.5, 39.9, 36.9, 29.3, 24.1, 23.6, 19.1,15.0, 8.2.

ESI-MS m/z: 767.2 (M−H₂O+H)⁺.

(+)-HR-ESI-TOF-MS m/z: 767.2794 [M−H₂O+H]⁺ (Calcd. for C₄₁H₄₃N₄O₉S767.2745).

Example 26

To a solution of compound 1 (621 mg, 1 mmol) in CH₃CN (100 mL, 0.01 M)was added compound 44-R (825 mg, 3 mmol) and cyanuric chloride (TCT)(248 mg, 40%). The reaction mixture was stirred at 85° C. for 66 h andthen aqueous saturated solution of NaHCO₃ was added and the mixture wasextracted with CH₂Cl₂. The combined organic layers were dried overanhydrous Na₂SO₄, filtered, and concentrated under vacuum. Flashchromatography (Hexane:EtOAc, from 9:1 to 1:9) gives compound 45-R (530mg, 58%).

R_(f)=0.4 (Hexane:EtOAc, 1:1).

¹H NMR (400 MHz, CDCl₃): δ 7.42-7.28 (m, 2H), 7.23-7.08 (m, 2H), 6.60(s, 1H), 6.20 (d, J=1.4 Hz, 1H), 6.04 (d, J=1.4 Hz, 1H), 6.01-5.92 (m,1H), 5.77 (s, 1H), 5.44-5.20 (m, 2H), 5.09 (s, 1H), 5.04-4.96 (m, 1H),4.71-4.55 (m, 2H), 4.34 (s, 1H), 4.30-4.18 (m, 3H), 3.79 (s, 3H), 3.53(dd, J=10.2, 4.4 Hz, 1H), 3.46 (m, 2H), 3.50-3.40 (m, 1H), 3.03-2.87 (m,2H), 2.67 (d, J=15.0 Hz, 1H), 2.47 (dd, J=15.6, 3.7 Hz, 1H), 2.40-2.32(m, 2H), 2.30 (s, 3H), 2.29 (s, 3H), 2.19-2.12 (m, 2H), 2.16 (s, 3H),2.09 (s, 3H).

ESI-MS m/z: 878.3 (M+H)⁺.

To a solution of compound 45-R (552 mg, 0.63 mmol) in CH₂Cl₂ (11.3 mL,18 mL/mmol) was added bis(triphenylphosphine)palladium(II) dichloride(70.7 mg, 0.1 mmol) and acetic acid (0.36 mL, 6.3 mmol). Tributyltinhydride (1.02 mL, 3.8 mmol) was added at 0° C. and the reaction mixturewas stirred at 0° C. for 0.5 h, and concentrated under vacuum The crudeobtained was diluted with EtOAc, saturated aqueous solution of NH₄Cl wasadded and the mixture was extracted with EtOAc. The combined organiclayers were dried over anhydrous Na₂SO₄, filtered, and concentratedunder vacuum. Flash chromatography (Hexane:EtOAc, from 100:1 to 1:100and EtOAc:CH₃OH, from 100:1 to 1:100) to afford compound 46-R (423 mg,85%).

R_(f)=0.3 (CH₂Cl₂:CH₃OH, 1:1).

¹H NMR (400 MHz, CDCl₃): δ 7.45-7.28 (m, 2H), 7.23-7.08 (m, 2H), 6.56(s, 1H), 6.19 (d, J=1.4 Hz, 1H), 6.05 (d, J=1.4 Hz, 1H), 4.98 (d, J=11.5Hz, 1H), 4.59 (s, 1H), 4.34 (s, 1H), 4.27 (dd, J=5.1, 1.7 Hz, 1H),4.22-4.16 (m, 2H), 3.80 (s, 3H), 3.49-3.39 (m, 2H), 3.31 (dq, J=9.8,5.5, 4.5 Hz, 2H), 2.95 (s, 1H), 2.83 (d, J=5.6 Hz, 2H), 2.74-2.51 (m,3H), 2.35 (s, 3H), 2.32-2.21 (m, 2H), 2.26 (s, 3H); 2.16 (s, 3H), 2.06(s, 3H).

ESI-MS m/z: 794.3 (M+H)⁺.

To a solution of compound 46-R (412 mg, 0.52 mmol) in CH₃CN:H₂O (1.39:1,36 mL, 0.015 M) was added AgNO₃ (1.76 g, 10.4 mmol). After 22 h at 23°C., the reaction was quenched with a mixture 1:1 of saturated aqueoussolutions of NaCl and NaHCO₃, stirred for 15 min, diluted with CH₂Cl₂,stirred for 5 min, and extracted with CH₂Cl₂. The combined organiclayers were dried over anhydrous Na₂SO₄, filtered, and concentratedunder vacuum. The residue obtained was purified by flash chromatography(CH₂Cl₂:CH₃OH, from 99:1 to 85:15) to give compound 47-R (175 mg, 43%).

R_(f)=0.1 (CH₂Cl₂:CH₃OH, 9:1).

¹H NMR (500 MHz, CDCl₃): δ 7.34 (dd, J=11.1, 7.9 Hz, 2H), 7.22-7.07 (m,2H), 6.57 (s, 1H), 6.17 (d, J=1.2 Hz, 1H), 6.01 (d, J=1.2 Hz, 1H), 5.11(d, J=11.2 Hz, 1H), 4.84 (s, 1H), 4.53-4.47 (m, 2H), 4.21-4.07 (m, 2H),3.80 (s, 3H), 3.56 (d, J=5.1 Hz, 1H), 3.43 (s, 1H), 3.24 (d, J=9.1 Hz,1H), 2.98-2.78 (m, 4H), 2.72-2.58 (m, 2H), 2.38 (s, 3H), 2.35-2.27 (m,2H), 2.28 (s, 3H), 2.14 (s, 3H), 2.08 (s, 3H).

¹³C NMR (101 MHz, CD₃OD): δ 170.6, 169.1, 155.0, 148.8, 145.6, 143.7,141.1, 140.8, 130.9, 129.7, 126.9, 124.2, 122.4, 121.1, 119.6, 118.9,118.7, 115.0, 113.2, 112.5, 111.0, 102.1, 91.3, 63.3, 60.4, 59.0, 58.4,55.3, 54.6, 52.6, 51.1, 44.9, 42.4, 39.8, 38.7, 29.4, 24.0, 23.2, 19.1,15.0, 8.3.

ESI-MS m/z: 767.2 (M−H₂O+H)⁺.

(+)-HR-ESI-TOF-MS m/z: 767.2806 [M−H₂O+H]⁺ (Calcd. for C₄₁H₄₃N₄O₉S767.2745).

Example 27. In Vitro Bioassays for the Detection of Antitumor Activity

The aim of this assay is to evaluate the in vitro cytostatic (ability todelay or arrest tumor cell growth) or cytotoxic (ability to kill tumorcells) activity of the samples being tested.

CELL LINES Name N^(o) ATCC Species Tissue Characteristics A549 CCL-185human lung lung carcinoma (NSCLC) HT29 HTB-38 human colon colorectaladenocarcinoma MDA- HTB-26 human breast breast adenocarcinoma MB- 231PSN1 CRM-CRL- human pancreas pancreas adenocarcinoma 3211 PC-3 CRL-1435human prostate prostate adenocarcinoma 22Rv1 CRL-2505 human prostateprostate carcinoma

Evaluation of Cytotoxic Activity Using the SBR and the MTT ColorimetricAssays

A colorimetric assay, using sulforhodamine B (SRB) reaction has beenadapted to provide a quantitative measurement of cell growth andviability (following the technique described by Skehan et al. J. Natl.Cancer Inst. 1990, 82, 1107-1112). Another colorimetric assay based on3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT)reduction to a purple formazan has been also used to assess theantiproliferative activity (following the technique described by Mosmannet al. J. Immunol. Meth. 1983, 65, 55-63).

These forms of assays employ 96-well cell culture microplates followingthe standards of the American National Standards Institute and theSociety for Laboratory Automation and Screening (ANSI SLAS 1-2004(R2012) Oct. 12, 2011. All the cell lines used in this study wereobtained from the American Type Culture Collection (ATCC) and derivefrom different types of human cancer.

A549, HT29, MDA-MB-231 and PSN1 cells were maintained in Dulbecco'sModified Eagle Medium (DMEM) while PC-3 and 22Rv1 cells were maintainedin Roswell Park Memorial Institute Medium (RPMI). All cell lines weresupplemented with 10% Fetal Bovine Serum (FBS), 2 mM L-glutamine, 100U/mL penicillin, and 100 U/mL streptomycin at 37° C., 5% CO₂ and 98%humidity. For the experiments, cells were harvested from subconfluentcultures using trypsinization and resuspended in fresh medium beforecounting and plating.

A549, HT29, MDA-MB-231 and PSN1 cells were seeded in 96 well microtiterplates, at 5000 cells per well in aliquots of 150 μL, and allowed toattach to the plate surface for 18 hours (overnight) in drug freemedium. After that, one control (untreated) plate of each cell line wasfixed (as described below) and used for time zero reference value.Culture plates were then treated with test compounds (50 μL aliquots of4× stock solutions in complete culture medium plus 4% DMSO) using ten2/5 serial dilutions (concentrations ranging from 10 to 0.003 μg/mL) andtriplicate cultures (1% final concentration in DMSO). After 72 hourstreatment, the antitumor effect was measured by using the SRBmethodology: Briefly, cells were washed twice with PBS, fixed for 15 minin 1% glutaraldehyde solution at room temperature, rinsed twice in PBS,and stained in 0.4% SRB solution for 30 min at room temperature. Cellswere then rinsed several times with 1% acetic acid solution andair-dried at room temperature. SRB was then extracted in 10 mM trizmabase solution and the absorbance measured in an automatedspectrophotometric plate reader at 490 nm.

An appropriate number of PC-3 and 22Rv1 cells, to reach a final celldensity in the assay ranging from 5,000 to 15,000 cells per welldepending on the cell line, were seeded in 96-well plates and allowed tostand in culture medium for 24 h at 37° C. under 5% CO2 and 98%humidity. Then, compounds or DMSO in culture medium were added to reacha final volume of 200 μL and the intended compound concentration in arange covering ten serial 2/5 dilutions starting from 0.1 μg/mL in 1%(v/v) DMSO. At this point a set of “time zero control plates” treatedwith 1% (v/v) DMSO were processed with MTT as described below. The restof the plates were incubated during 72 h under the aforementionedenvironmental conditions. Afterwards 50 μL of a 1 mg/mL MTT solution inculture medium were added to the wells and incubated for 6-8 hours at37° C. to allow formazan crystals generation. Culture medium was thenremoved and 100 μL of neat DMSO added to each well to dissolve theformazan product into a coloured solution whose absorbance at 540 nm wasfinally measured in a PolarStar Omega microplate multilabel reader (BMGLabtech, Ortenberg, Germany).

Effects on cell growth and survival were estimated by applying the NCIalgorithm (Boyd M R and Paull K D. Drug Dev. Res. 1995, 34, 91-104). Thevalues obtained in triplicate cultures were fitted by nonlinearregression to a four-parameters logistic curve by nonlinear regressionanalysis. Three reference parameters were calculated (according to theaforementioned NCI algorithm) by automatic interpolation of the curvesobtained by such fitting: GI₅₀=compound concentration that produces 50%cell growth inhibition, as compared to control cultures; TGI=total cellgrowth inhibition (cytostatic effect), as compared to control cultures,and LC₅₀=compound concentration that produces 50% net cell killingcytotoxic effect).

Tables 1-7 illustrate data on the biological activity of compounds ofthe present invention together with biological activity of the referencecompounds. Tables 8-9 provide data on the biological activity of severalcompounds of the invention compared to their analogues with a carboxylicacid group. Compounds A, B, E, F, ET-736, PM01183, 14-S, 15-S, 28-S,28-R, 29-S, and 29-R, are not part of the present invention.

TABLE 1 Biological activity (Molar)

                 

MDA- MDA-MB- A549 HT29 MB-231 PSN1 PC-3 22Rv1 A549 HT29 231 PSN1 GI₅₀ 3-4.03E−10 2.77E−10 4.91E−10   9.95E−10 A   8.36E−09   7.71E−09 7.07E−091.29E−08 TGI S 6.17E−10 >1.26E−07   5.29E−10   1.64E−09   8.87E−09  8.36E−09 9.38E−09 1.54E−08 LC₅₀ >1.26E−07   >1.26E−07  6.17E−10 >1.26E−07 >1.29E−07 >1.29E−07 1.41E−08 1.93E−08 GI₅₀ 3a-3.11E−09 2.99E−09 2.87E−09   2.15E−09 TGI S 3.23E−09 3.23E−09 3.59E−09  3.59E−09 LC₅₀ >1.20E−07   >1.20E−07   4.90E−09   1.20E−08 GI₅₀ 10-2.05E−08 1.14E−08 4.79E−09   7.64E−09 TGI S 3.08E−08 1.25E−08 8.44E−09  1.25E−08 LC₅₀ 7.53E−08 >1.14E−06   1.60E−08   2.39E−08 GI₅₀ 11-8.45E−09 3.41E−09 2.27E−09   3.28E−09 TGI S 2.65E−08 >1.26E−07  3.41E−09   4.54E−09 LC₅₀ >1.26E−07   >1.26E−07   6.43E−09   8.07E−09GI₅₀ 4- 1.27E−09 1.27E−09 1.22E−09   1.78E−09 8.08E−10 3.58E−10 C  2.73E−08   2.08E−08 2.60E−08 3.64E−08 TGI S 1.40E−09 1.40E−09 2.55E−09  2.29E−09   6.63E−08   2.34E−08 5.46E−08 4.42E−08LC₅₀ >1.27E−07   >1.27E−07   6.50E−09  3.44E−09 >1.30E−07 >1.30E−07 >1.30E−07   6.50E−08 GI₅₀ 4a- 3.99E−093.14E−09 3.39E−09   3.02E−09 TGI S 6.17E−09 3.39E−09 5.44E−09   3.27E−09LC₅₀ >1.21E−07   >1.21E−07   1.00E−08   3.51E−09 GI₅₀ 12- 2.04E−084.85E−09 5.23E−09   3.44E−09 TGI S 5.61E−08 8.42E−09 8.42E−09   5.49E−09LC₅₀ >1.28E−07   >1.28E−07   1.53E−08   1.21E−08 GI₅₀ 13- 1.15E−081.15E−08 1.15E−08   1.96E−08 TGI S 1.61E−08 1.27E−08 1.27E−08   2.88E−08LC₅₀ 2.42E−08 >1.15E−06   1.38E−08   4.61E−08

TABLE 2 Biological activity (Molar)

           

MDA-MB- MDA-MB- A549 HT29 231 PSN1 A549 HT29 231 PSN1 GI₅₀ 3-R 4.03E−102.77E−10 2.77E−10 3.90E−10 B 2.06E−08 8.48E−09 9.00E−09 1.93E−08 TGI5.79E−10 >1.26E−07   5.04E−10 6.05E−10 2.19E−08 9.13E−09 1.67E−082.06E−08 LC₅₀ >1.26E−07   >1.26E−07  1.25E−09 >1.26E−07   >1.29E−07   >1.29E−07   3.47E−08 2.31E−08 GI₅₀ 10-R3.76E−09 3.08E−09 2.85E−09 2.62E−09 TGI 5.93E−09 >1.14E−07   4.33E−093.88E−09 LC₅₀ >1.14E−07   >1.14E−07   7.18E−09 6.61E−09 GI₅₀ 11-R1.77E−09 1.39E−09 1.01E−09 1.39E−09 TGI 4.54E−09 >1.26E−07   1.51E−091.89E−09 LC₅₀ >1.26E−07   >1.26E−07   2.65E−09 >1.26E−07   GI₅₀ 4-R1.27E−09 1.26E−09 1.27E−09 4.59E−10 D 1.25E−08 1.03E−08 9.88E−092.08E−08 TGI 1.40E−09 1.40E−09 1.40E−09 8.54E−10 2.86E−08 2.34E−081.95E−08 2.21E−08 LC₅₀ >1.27E−07   >1.27E−07   1.53E−092.55E−09 >1.30E−07   >1.30E−07   5.33E−08 2.47E−08 GI₅₀ 12-R 1.40E−095.74E−10 3.19E−10 4.98E−10 TGI 2.93E−09 1.10E−09 6.76E−10 1.22E−09 LC₅₀1.22E−08 2.93E−09 1.40E−09 >1.28E−07   GI₅₀ 13-R 7.26E−09 6.91E−094.95E−09 2.88E−09 TGI 7.72E−09 7.60E−09 7.95E−09 3.11E−09LC₅₀ >1.15E−07   >1.15E−07   1.38E−08 3.46E−09

TABLE 3 Biological activity (Molar)

       

MDA-MB- MDA-MB- A549 HT29 231 PSN1 PC-3 22Rv1 A549 HT29 231 PSN1 GI₅₀38-S 8.05E−09 4.53E−09 2.52E−09 5.03E−09 A 8.36E−09 7.71E−09 7.07E−091.29E−08 TGI 8.55E−09 7.05E−09 4.28E−09 8.18E−09 8.87E−09 8.36E−099.38E−09 1.54E−08 LC₅₀ 9.44E−09 >1.26E−07   7.80E−091.51E−08 >1.29E−07   >1.29E−07   1.41E−08 1.93E−08 GI₅₀ 45-S 1.82E−081.82E−08 1.71E−08 1.94E−08 TGI 1.94E−08 1.94E−08 2.16E−08 2.62E−08 LC₅₀2.16E−08 >1.14E−07   2.96E−08 3.64E−08 GI₅₀ 46-S 8.19E−09 2.77E−093.65E−09 3.15E−09 TGI 2.14E−08 6.17E−09 6.80E−09 4.79E−09LC₅₀ >1.26E−07   >1.26E−07   1.26E−08 9.20E−09 GI₅₀ 39-S 4.84E−093.94E−09 3.44E−09 8.02E−09 2.78E−09 4.81E−10 C 2.73E−08 2.08E−082.60E−08 3.64E−08 TGI 8.27E−09 6.74E−09 7.13E−09 1.02E−08 6.63E−082.34E−08 5.46E−08 4.42E−08 LC₅₀ 1.65E−08 >1.27E−07   1.78E−081.27E−08 >1.30E−07   >1.30E−07   >1.30E−07   6.50E−08 GI₅₀ 47-S 1.40E−084.33E−09 6.24E−09 5.99E−09 TGI 2.80E−08 6.75E−09 9.68E−09 8.54E−09LC₅₀ >1.27E−07   >1.27E−07   1.66E−08 1.27E−08

TABLE 4 Biological activity (Molar)

       

MDA-MB- MDA-MB- A549 HT29 231 PSN1 A549 HT29 231 PSN1 GI₅₀ 38-R 6.54E−105.41E−10 4.53E−10 6.54E−10 B 2.06E−08 8.48E−09 9.00E−09 1.93E−08 TGI1.04E−09 5.91E−10 8.43E−10 9.94E−10 2.19E−08 9.13E−09 1.67E−08 2.06E−08LC₅₀ >1.26E−07   >1.26E−07   2.01E−09 1.76E−09 >1.29E−07   >1.29E−07  3.47E−08 2.31E−08 GI₅₀ 45-R 1.82E−08 1.25E−08 9.57E−09 1.06E−08 TGI1.94E−08 2.28E−08 1.94E−08 1.94E−08 LC₅₀ 2.39E−08 >1.14E−07   4.33E−083.76E−08 GI₅₀ 46-R 1.51E−09 1.21E−09 1.23E−09 9.95E−10 TGI 2.77E−091.39E−09 1.39E−09 1.51E−09 LC₅₀ >1.26E−07   >1.26E−07   1.51E−092.65E−09 GI₅₀ 39-R 2.67E−10 2.93E−10 2.04E−10 3.65E−10 D 1.25E−081.03E−08 9.88E−09 2.08E−08 TGI 4.33E−10 6.24E−10 5.98E−10 5.73E−102.86E−08 2.34E−08 1.95E−08 2.21E−08 LC₅₀ >1.27E−07   >1.27E−07  2.80E−09 1.06E−09 >1.30E−07   >1.30E−07   5.33E−08 2.47E−08 GI₅₀ 47-R2.04E−09 8.03E−10 5.99E−10 1.40E−09 TGI 3.82E−09 1.40E−09 1.17E−092.04E−09 LC₅₀ 1.40E−08 >1.27E−07   2.55E−09 3.31E−09

TABLE 5 Biological activity (Molar)

       

MDA-MB- MDA-MB- A549 HT29 231 PSN1 A549 HT29 231 PSN1 GI₅₀ 18-S 1.70E−091.21E−09 1.21E−09 9.59E−10 E 3.28E−09 3.15E−09 2.27E−09 2.77E−09 TGI3.03E−09 1.34E−09 1.34E−09 1.34E−09 3.40E−09 3.40E−09 3.78E−09 4.53E−09LC₅₀ >1.21E−07   >1.21E−07   1.58E−09 >1.21E−07   4.41E−09 >1.26E−07  7.43E−09 8.94E−09 GI₅₀ 25-S 7.17E−09 7.17E−09 5.84E−09 6.84E−09 TGI7.61E−09 7.72E−09 9.04E−09 9.26E−09 LC₅₀ >1.10E−07   >1.10E−07  1.54E−08 1.43E−08 GI₅₀ 26-S 1.12E−08 2.79E−09 1.34E−09 3.04E−09 TGI2.19E−08 3.16E−09 1.94E−09 3.28E−09 LC₅₀ >1.22E−07   >1.22E−07  3.89E−09 3.52E−09 GI₅₀ 19-S 3.07E−09 1.35E−09 1.96E−09 2.95E−09 PM011833.31E−09 1.91E−09 2.29E−09 3.19E−09 TGI 3.31E−09 1.60E−09 3.31E−093.19E−09 3.57E−09 4.46E−09 3.95E−09 3.95E−09LC₅₀ >1.23E−07   >1.23E−07  1.10E−08 >1.23E−07   >1.27E−07   >1.27E−07   1.02E−08 5.73E−09 GI₅₀ 27-S6.02E−09 1.23E−09 1.19E−09 1.97E−09 TGI 1.12E−08 1.35E−09 1.23E−092.83E−09 LC₅₀ >1.23E−07   >1.23E−07   1.35E−09 4.55E−09

TABLE 6 Biological activity (Molar)

       

MDA-MB- MDA-MB- A549 HT29 231 PSN1 A549 HT29 231 PSN1 GI₅₀ 18-R 1.21E−091.21E−09 1.21E−09 5.70E−10 E 3.28E−09 3.15E−09 2.27E−09 2.77E−09 TGI1.34E−09 1.34E−09 1.34E−09 1.06E−09 3.40E−09 3.40E−09 3.78E−09 4.53E−09LC₅₀ >1.21E−07   >1.21E−07   1.46E−09 >1.21E−07   4.41E−09 >1.26E−07  7.43E−09 8.94E−09 GI₅₀ 25-R 1.32E−09 1.54E−09 1.21E−09 1.21E−09 TGI2.43E−09 2.76E−09 2.54E−09 2.32E−09 LC₅₀ 9.92E−09 >1.10E−07   8.38E−096.73E−09 GI₅₀ 26-R 1.94E−09 7.29E−10 1.17E−09 9.72E−10 TGI 3.40E−091.58E−09 1.22E−09 1.70E−09 LC₅₀ >1.22E−07   >1.22E−07   1.46E−093.52E−09 GI₅₀ 19-R 1.47E−09 1.72E−09 1.23E−09 1.23E−09 PMO 3.31E−091.91E−09 2.29E−09 3.19E−09 TGI 3.56E−09 1.72E−09 1.35E−09 1.35E−09 11833.57E−09 4.46E−09 3.95E−09 3.95E−09LC₅₀ >1.23E−07   >1.23E−07   >1.23E−07  1.47E−09 >1.27E−07   >1.27E−07   1.02E−08 5.73E−09 GI₅₀ 27-R 2.09E−095.04E−10 3.07E−10 6.39E−10 TGI 3.93E−09 5.53E−10 5.41E−10 1.17E−09 LC₅₀1.01E−08 >1.23E−07   8.60E−10 2.46E−09

TABLE 7 Biological activity (Molar)

MDA-MB- MDA-MB- A549 HT29 231 PSN1 A549 HT29 231 PSN1 GI₅₀ 31 1.96E−081.05E−08 8.89E−09 6.80E−09 F 3.80E−08 2.09E−08 1.96E−08 3.27E−08 TGI2.09E−08 1.57E−08 1.70E−08 1.57E−08 7.20E−08 2.36E−08 3.40E−08 6.02E−08LC₅₀ 2.35E−08 >1.31E−07   3.53E−08 4.31E−08 >1.31E−07   >1.31E−07  7.33E−08 1.07E−07 GI₅₀ 32 6.88E−09 6.88E−09 4.76E−09 6.09E−09 ET-7362.25E−08 2.12E−08 2.12E−08 3.97E−08 TGI >1.32E−08   >1.32E−08   1.05E−088.34E−09 4.77E−08 2.25E−08 2.52E−08 5.96E−08LC₅₀ >1.32E−08   >1.32E−08   >1.32E−08  1.20E−08 >1.32E−07   >1.32E−07   4.77E−08 1.02E−07 GI₅₀ 34 5.91E−085.41E−08 4.53E−08 5.41E−08 E 3.28E−09 3.15E−09 2.27E−09 2.77E−09 TGI8.05E−08 8.55E−08 7.67E−08 5.91E−08 3.40E−09 3.40E−09 3.78E−09 4.53E−09LC₅₀ >1.26E−07   1.25E−07 1.12E−07 >1.26E−07   4.41E−09 >1.26E−07  7.43E−09 8.94E−09 GI₅₀ 35 8.14E−09 7.89E−09 4.58E−09 6.24E−09 PM011833.31E−09 1.91E−09 2.29E−09 3.19E−09 TGI 8.78E−09 8.65E−09 8.27E−099.03E−09 3.57E−09 4.46E−09 3.95E−09 3.95E−09LC₅₀ >1.27E−07   >1.27E−07   1.65E−08 1.40E−08 >1.27E−07   >1.27E−07  1.02E−08 5.73E−09

TABLE 8 Biological activity (Molar)  

MDA-MB- MDA-MB- A549 HT29 231 PSN1 A549 HT29 231 PSN1 GI₅₀ 3-S 4.03E−102.77E−10 4.91E−10 9.95E−10 14-S >1.24E−07 1.21E−07 5.45E−08 >1.24E−07  TGI 6.17E−10 >1.26E−07   5.29E−10 1.64E−09 >1.24E−07 >1.24E−07  1.13E−07 >1.24E−07   LC₅₀ >1.26E−07   >1.26E−07  6.17E−10 >1.26E−07   >1.24E−07 >1.24E−07   >1.24E−07   >1.24E−07   GI₅₀4-S 1.27E−09 1.27E−09 1.22E−09 1.78E−09 15-S >1.25E−06 3.00E−07 1.63E−072.38E−07 TGI 1.40E−09 1.40E−09 2.55E−09 2.29E−09 >1.25E−06 5.13E−072.13E−07 4.63E−07 LC₅₀ >1.27E−07   >1.27E−07   6.50E−093.44E−09 >1.25E−06 9.14E−07 2.75E−07 8.39E−07 GI₅₀ 18-S 1.70E−091.21E−09 1.21E−09 9.59E−10 28-S   4.89E−07 2.51E−07 1.67E−07 2.51E−07TGI 3.03E−09 1.34E−09 1.34E−09 1.34E−09 >1.19E−06 3.46E−07 2.51E−073.94E−07 LC₅₀ >1.21E−07   >1.21E−07   1.58E−09 >1.21E−07   >1.19E−066.33E−07 3.94E−07 6.92E−07 GI₅₀ 19-S 3.07E−09 1.35E−09 1.96E−09 2.95E−0929-S   6.15E−07 3.62E−07 2.17E−07 3.86E−07 TGI 3.31E−09 1.60E−093.31E−09 3.19E−09 >1.21E−06 5.31E−07 3.74E−07 5.07E−07LC₅₀ >1.23E−07   >1.23E−07   1.10E−08 >1.23E−07   >1.21E−06 8.32E−076.88E−07 6.88E−07

TABLE 9 Biological activity (Molar)

MDA-MB- MDA-MB- A549 HT29 231 PSN1 A549 HT29 231 PSN1 GI50 18-R 1.21E−091.21E−09 1.21E−09 5.71E−10 28-R 1.67E−07 3.10E−08 1.91E−08 2.15E−08 TGI1.34E−09 1.34E−09 1.34E−09 1.06E−09 3.58E−07 3.34E−08 3.22E−08 3.58E−08LC50 >1.21E−07   >1.21E−07  1.46E−09 >1.21E−07   >1.19E−06   >1.19E−06   9.19E−08 6.68E−08 GI50 19-R1.47E−09 1.72E−09 1.23E−09 1.23E−09 29-R 9.05E−08 3.02E−08 1.69E−083.02E−08 TGI 3.56E−09 1.72E−09 1.35E−09 1.35E−09 1.93E−07 3.26E−082.77E−08 3.14E−08 LC50 >1.23E−07   >1.23E−07   >1.23E−07  1.47E−09 >1.21E−06   >1.21E−06   1.57E−07 3.50E−08

The compounds of the present invention are shown to have high potency invitro, when compared against reference compounds. This demonstrates thatthe compounds according to the present invention exhibit high cytoxiticytowards cancer cells and are useful in the treatment of cancer.

Example 28. MTD and MTMD Determination

Female CD-1 or Athymic Nude-Fox1 nu/nu mice (Envigo) were utilized forall experiments. Animals (N=10/cage) were housed in individuallyventilated cages (Sealsafe Plus®, Techniplast S.P.A.), on a 12-hourlight-dark cycle at 21-23° C. and 40-60% humidity. Mice were allowedfree access to irradiated standard rodent diet (Tecklad 2914C) andsterilized water. Animals were acclimated for five days prior to beingindividually tattoo-identified. Animal protocols were reviewed andapproved according to the regional Institutional Animal Care and UseCommittees.

Mice were randomly allocated into experimental groups and intravenouslyadministered, once for the MTD (Maximum Tolerated Dose) determination orone administration a week during three consecutive weeks, for the MTMD(Maximum Tolerated Multiple Dose) determination study. The animals wereadministered with white formulation or with compound dissolved in theexperimental formulation at different concentrations. The volumeadministered was always 10 mL/kg. Once administered, animals weremonitored for clinical signs of systemic toxicity, changes in bodyweight and mortality up to 14 days after the administration.

MTD results are summarized in Table 10

TABLE 10 Route/ MTD Compound Schedule Doses (mg/Kg) (mg/kg)  4-S iv/SD0.00, 0.25, 0.50, 1.00, 1.50, 1.0 2.00, 2.50, 5.00  4-R 0.25 19-S 0.519-R 0.00, 0.10, 0.15, 0.25, 0.50, 1.00, 0.15 1.50, 2.00, 2.50, 5.00Comp C 0.00, 0.25, 0.50, 1.00, 1.50, 2.00, 3.0 2.50, 3.00, 4.00, 5.00Comp D 0.00, 0.25, 0.50, 1.00, 2.00, 4.00, 0.5 6.00, 8.00 32 0.00, 0.25,0.50, 1.00, 1.50, 2.00, 0.5 2.50, 5.00

MTMD results are summarized in Table 11

TABLE 11 Route/ MTMD Compound Schedule Doses (mg/Kg) (mg/kg)  4-S iv/Q7d× 3 0.00, 0.50, 0.75, 1.00, 1.25 1.25  4-R 0.00, 0.15, 0.20, 0.25, 0.300.30 12-S 0.00, 0.10, 0.25, 0.50, 0.75, 0.25 1.00, 1.25, 1.50, 2.00,2.50, 5.00 12-R 0.00, 0.010, 0.025, 0.050, 0.05 0.075, 0.10, 0.25, 0.50,0.75, 1.00, 1.25, 1.50, 2.00, 2.50, 5.00 19-S 0.00, 0.10, 0.25, 0.50,0.75 0.75 19-R 0.00, 0.025, 0.075, 0.10, 0.15 0.15 Comp C 0.0, 1.0, 1.5,2.0, 3.0, 4.0 3.0 Comp D 0.00, 0.10, 0.25, 0.50, 0.75 0.5 32 0.00, 0.10,0.25, 0.50, 0.75 0.5 35 0.00, 0.10, 0.25, 0.50, 0.75 0.25 39-S 0.00,0.01, 0.025, 0.05, 0.075, 1.25 0.10, 0.25, 0.50, 0.75, 1.00, 1.25, 1.50,2.00, 2.50, 5.00 47-R 0.00, 0.01, 0.025, 0.05, 0.075, 0.1 0.10, 0.25,0.50, 0.75, 1.00, 1.25, 1.50, 2.00, 2.50, 5.00 ET-736 0.00, 0.10, 0.25,0.50, 0.75 0.5 PM01183 0.00, 0.14, 0.18 0.18 iv, intravenously Q7d × 3,three cumulated doses administered in a weekly basis.

Examples 29-40. In Vivo Xenografts

Female athymic nu/nu mice (Harlan Laboratories Models, S. L. Barcelona,Spain or Envigo, Spain) were utilized for all experiments. Animal werehoused in individually ventilated cages Sealsafe® Plus, TechniplastS.P.A.), up to ten per cage on a 12-hour light-dark cycle at 21-23° C.and 40-60% humidity. Mice were allowed free access to irradiatedstandard rodent diet (Tecklad 2914C) and sterilized water. Animals wereacclimated for at least 5 days prior to tumor implantation with a tumorcell suspension.

CELL LINES Name N^(o) ATCC N^(o) ECCC* Species Tissue CharacteristicsHT1080 CCL-121 — human connective Fibrosarcoma MDA-MB- HTB-26 — humanbreast Breast 231 adenocarcinoma H460 HTB-177 — human lung, pleuralNSCLC effusion A2780 — 93112519 human ovarian Ovarian carcinoma HGC27 —94042256 human gastric Gastric carcinoma H526 CRL-5811 — human lung SCLCH82 HTB-175 — human lung SCLC PC3 CLR-1435 — human prostate; Prostaticderived from adenocarcinoma metastatic site: bone DU145 HTB-81 humanprostate; Prostatic carcinoma derived from metastatic site: brain 22Rv1CRL-2505 human prostate Prostatic carcinoma *European Collection of CellCultures

HT1080 cells were maintained in vitro at 37° C. with 5% CO₂ in MinimumEssential Medium Eagle (MEME) (Sigma-Aldrich, Co). Each animal wasorthotopically implanted into gastroecnemius muscle by an intramuscularinjection using a 26 G needle and a 1 cc syringe at 4-6 weeks of age,with 10×10⁶ HT1080 cells, suspended in serum free medium, withoutantibiotics.

MDA-MB-231 cells were maintained in vitro at 37° C. with 5% CO₂ inDulbecco's Modified Eagle's Medium (Sigma-Aldrich, Co). Culture cellswere passaged every 3 to 5 days upon reaching confluence. Each animalwas subcutaneously implanted (on the right flank using 26 G needle and a1 cc syringe) at 4-6 weeks of age with 7.5×10⁶ MDA-MB-231 cellssuspended in 0.05 mL of a solution consisting of 50% Matrigel® (CorningIncorporated Life Sciences) and 50% medium without serum or antibiotics.

H460 cells were maintained in vitro at 37° C. with 5% CO₂ in Dulbecco'sModified Eagle's Medium (Sigma-Aldrich, Co). Culture cells were passagedevery 3 to 5 days upon reaching confluence. Each animal wassubcutaneously implanted (on the right flank using 26 G needle and a 1cc syringe) at 4-6 weeks of age with 5×10⁶ H460 cells suspended in 0.05mL of a solution consisting of 50% Matrigel® (Corning Incorporated LifeSciences) and 50% medium without serum or antibiotics.

A2780 cells were maintained in vitro at 37° C. with 5% CO₂ in RPMI-1640(Sigma-Aldrich, Co). Culture cells were passaged every 3 to 5 days uponreaching confluence. Each animal was subcutaneously implanted (on theright flank using 26 G needle and a 1 cc syringe) at 4-6 weeks of agewith 10×10⁶ A2780 cells suspended in 0.05 mL of a solution consisting of50% Matrigel® (Corning Incorporated Life Sciences) and 50% mediumwithout serum or antibiotics.

HGC27 cells were maintained in vitro at 37° C. with 5% CO₂ in Iscove'sModified Dulbecco's Medium (Sigma Aldrich, Co). Culture cells werepassage every 3 to 5 days on reaching confluence. Each animal wassubcutaneously implanted (on the right flank using 26 G needle and a 1cc syringe) at 4-6 weeks of age with 5×10⁶ HGC-27 cells suspended in0.05 mL of a solution consisting of 50% Matrigel® (Corning IncorporatedLife Sciences), 50% medium without serum or antibiotics.

H526 cells were maintained in vitro at 37° C. with 5% CO₂ in RPMI-1640Medium (Sigma-Aldrich, Co). H526 cells were grown as a suspension andmaintained by addition of fresh medium, as the cell density increases,every 2 to 3 days. Every week, culture was reestablished bycentrifugation of the suspension with subsequent resuspension in freshmedium at a concentration of 1×10⁵ cell/mL. Each animal wassubcutaneously implanted (on the right flank using 26 G needle and a 1cc syringe) at 4-6 weeks of age with 5×10⁶ H526 cells suspended in 0.05mL of a solution consisting of 50% Matrigel® (Corning Incorporated LifeSciences) and 50% medium without serum or antibiotics.

H82 cells were maintained in vitro at 37° C. with 5% CO₂ in RPMI-1640Medium (Sigma-Aldrich, Co). H82 cells were grown as a suspension andmaintained by addition of fresh medium, as the cell density increases,every 2 to 3 days. Every week, culture was reestablished bycentrifugation of the suspension with subsequent resuspension in freshmedium at a concentration of 1×10⁵ cell/ml. Animals were subcutaneouslyimplanted (on the right flank using 26 G needle and a 1 cc syringe) at4-6 weeks of age with 5×10⁶ H82 cells, suspended in 0.05 mL of asolution consisting of 50% Matrigel® (Corning Incorporated LifeSciences) and 50% medium without serum or antibiotics.

PC3 cells were maintained in vitro at 37° C. with 5% CO2 in RPMI-1640Medium (Sigma-Aldrich, Co). Culture cells were passaged every 3 to 5days upon reaching confluence. Each female athymic mice wassubcutaneously implanted (on the right flank using a 26 G needle and a 1cc syringe) at 4-6 weeks of age with 3×10⁶ PC3 cells suspended in 0.05mL of a solution consisting of 50% Matrigel® Matrix (CorningIncorporated Life Sciences) and 50% medium without serum or antibiotics.In this model, instead of male, female animals were used because PC-3growth is not hormone dependant.

DU-145 cells were maintained in vitro at 37° C. with 5% CO2 in RPMI-1640Medium (Sigma-Aldrich, Co). Culture cells were passaged every 3 to 5days upon reaching confluence Each male athymic mice was subcutaneouslyimplanted (on the right flank using a 26 G needle and a 1 cc syringe) at4-6 weeks of age with 5×10⁶ DU-145 cells suspended in 0.05 mL of asolution consisting of 50% Matrigel® Matrix (Corning Incorporated LifeSciences) and 50% medium without serum or antibiotics.

22Rv1 cells were maintained in vitro at 37° C. with 5% CO₂ in RPMI-1640Medium (Sigma-Aldrich, Co). Culture cells were passage every 3 to 5 daysupon reaching confluence. Each male athymic mice was subcutaneouslyimplanted (on the right flank using 26 G needle and a 1 cc syringe) at4-6 weeks of age with 5×10⁶ 22 Rv1 cells suspended in 0.05 mL of asolution consisting of 50% Matrigel® Matrix (Corning Incorporated LifeSciences) and 50% medium without serum or antibiotics.

Treatment tolerability was assessed by monitoring body weight evolution,clinical signs of systemic toxicity, as well as evidences of localdamage in the injection site.

In xenograft studies with HT1080 cell line:

-   -   Total diameter (tumor+leg) measurements were determined by using        digital caliper (Fowler Sylvac, S235PAT). This total diameter        and animal body weights were measured 2-3 times per week        starting from the first day of treatment (day 0).    -   When total diameter reached a length of about 7.0-8.0 mm, mice        were randomly allocated into the treatments and control groups        (N=8-10/group) based on body weight and tumor measurements by        using NewLab Oncology Software (version 2.25.06.00).    -   Comparison of the median total diameter (tumor+leg) in the        treatment groups to the median total diameter (tumor+leg) in the        control group was used for evaluation of the antitumoral        efficacy.    -   Animals were euthanized when their total leg diameter reached        ca. 18 mm.

In xenograft studies with other cell lines:

-   -   Tumor volume was calculated using the equation (a·b²)/2, where        a: length (longest diameter) and b: width (shortest diameter)        were measured in mm by using digital caliper (Fowler Sylvac,        S235PAT). Tumor dimensions and body weights were recorded 2-3        times per week starting from the first day of treatment.    -   When tumors reached ca. 150-250 mm³, tumor bearing animals        (N=8-10/group) were randomly allocated into the treatment        groups, based on body weight and tumor measurements by using        NewLab Oncology Software (version 2.25.06.00).    -   Comparison between median tumor volume of treated groups and        control group was used for evaluation of the antitumoral        efficacy.    -   Animals were euthanized when their tumors reached ca. 2000 mm³        and/or severe necrosis was seen.

Treatments producing >20% lethality and/or 20% net body weight loss wereconsidered toxic.

Tables and figures summarize the data obtained from completeexperimental groups, i.e. those groups keeping the initial number ofanimals, n=8-10. However, once the first animal is sacrificed due to atumor length >18 mm or a tumor size >2000 mm³, the experimental groupwill be considered incomplete. Therefore, data generated subsequently tothe sacrifice day and onwards will not be presented (i.e. neither intables nor in the figures).

Example 29. In Vivo Studies to Determine the Effect of 4-S and 12-S inSeveral Xenograft Models

4-S, 12-S and compound C were provided in the form of freeze-dried vialsof lyophilized product. Each vial was reconstituted with water forinfusion to a concentration of 0.5 mg/mL. Further dilutions were madewith 5% dextrose solution for injection to the dosing formulationconcentration. The administered doses of 4-S, 12-S and compound C were1.25 mg/kg, 0.25 mg/kg and 3.0 mg/kg, respectively.

Placebo was provided in the form of lyophilised cake containing 100 mgSucrose+Potassium dihydrogen phosphate 6.8 mg+Phosphoric acid q.s. pH3.8-4.5 which was reconstituted with water for infusion.

In these experiments, 4-S, 12-S and Compound C, as well as placebo, wereintravenously administered once per week for 3 consecutive weeks, onDays 0, 7 and 14, whenever it was possible.

Example 29a. In Vivo Studies to Determine the Effect of 4-S and 12-S inHuman Fibrosarcoma Xenografts

The aim of this study was to compare the antitumoral activity of 4-S and12-S with the antitumoral activity of compound C by using a xenograftmodel of human sarcoma.

The tumor model used in this study was HT1080 cell line.

Table 12 reports the total diameter (tumor+leg) evaluation of HT1080tumors in mice treated with placebo, compound C, 4-S, and 12-S. Theseresults are also showed in FIG. 1.

TABLE 12 Total diameter (tumor + leg) (mm) Days Control Compound C 4-S12-S 0.0 7.5 7.5 7.5 7.5 2.0 9.4 8.8 7.7 8.2 5.0 11.4 9.0 8.3 8.6 7.012.1 9.6 8.8 9.5 9.0 13.2 10.2 8.4 10.0 12.0 14.5 10.2 8.4 11.2 14.015.2 11.2 9.6 11.7 16.0 15.9 12.4 10.0 12.7 19.0 18.0 13.3 10.4 13.521.0 15.2 12.1 14.4 23.0 18.0 12.7 16.5 27.0 13.5 15.2 30.0 15.6 16.433.0 18.0

Example 29b. In Vivo Studies to Determine the Effect of 4-S and 12-S inHuman Breast Xenografts

The aim of this study was to compare the antitumoral activity of 4-S and12-S with the antitumoral activity of compound C by using a xenograftmodel of human breast cancer.

The tumor model used in this study was MDA-MB-231 cell line.

Table 13 reports the median tumor volume evaluation of MDA-MB-231 tumorsin mice treated with placebo, compound C, 4-S, and 12-S. These resultsare also showed in FIG. 2.

TABLE 13 Median Tumor Volume (mm³) Days Control Compound C 4-S 12-S 0.0149.4 149.4 150.6 150.2 2.0 240.0 217.1 197.3 229.9 5.0 325.1 281.3250.9 290.5 7.0 407.8 338.6 265.0 398.2 9.0 514.8 385.1 272.5 508.9 12.0648.1 400.4 270.6 602.5 14.0 799.0 436.9 281.3 751.0 16.0 1002.5 585.7293.6 977.7 19.0 1233.9 774.7 322.1 1252.6 21.0 1539.1 965.9 324.41560.7 23.0 2006.5 1215.2 326.6 2005.9 26.0 2027.7 1503.2 398.8 2066.228.0 1785.3 501.8 30.0 2037.1 654.8 33.0 856.7 35.0 1147.1 37.0 1635.9

Example 29c. In Vivo Studies to Determine the Effect of 4-S and 12-S inHuman Lung Tumor Xenografts

The aim of this study was to compare the antitumoral activity of 4-S and12-S with the antitumoral activity of compound C by using threedifferent xenograft models of human lung cancer. These models correspondto non-small cell lung cancer (H-460 cell line) and to small cell lungcancer (H526 and H82 cell lines).

Table 14 reports the median tumor volume evaluation of H460 tumors inmice treated with placebo, compound C, 4-S, and 12-S. These results arealso showed in FIG. 3.

TABLE 14 Median Tumor Volume (mm³) Days Control Compound C 4-S 12-S 0.0187.4 186.1 185.9 186.0 2.0 577.5 395.4 310.9 460.5 5.0 1352.0 665.9634.6 922.4 7.0 1642.9 929.5 959.1 1252.1 9.0 2025.0 1063.7 1064.91409.4 12.0 1436.5 1421.0 1531.7 14.0 2025.0 1845.5 2025.0 16.0 2025.02025.0

Table 15 reports the median tumor volume evaluation of H526 tumors inmice treated with placebo, compound C, 4-S and 12-S. These results arealso showed in FIG. 4.

TABLE 15 Median Tumor Volume (mm³) Days Control Compound C 4-S 12-S 0.0217.2 217.9 211.8 212.7 2.0 410.7 262.4 279.0 412.7 4.0 778.5 108.3 98.8637.9 7.0 1083.2 129.8 56.7 968.5 9.0 1371.0 85.9 62.5 1250.3 11.01782.0 52.3 32.0 1568.0 14.0 2025.0 54.1 18.0 2025.0 16.0 47.3 32.0 21.04.0 4.0 28.0 4.0 4.0 35.0 4.0 4.0 42.0 62.5 4.0 49.0 53.5 4.0

Table 16 reports the median tumor volume evaluation of H82 tumors inmice treated with placebo, compound C, 4-S and 12-S. These results arealso showed in FIG. 5.

TABLE 16 Median Tumor Volume (mm³) Days Control Compound C 4-S 12-S 0.0171.6 170.5 168.3 174.0 2.0 439.4 265.3 215.2 360.1 5.0 1024.7 488.7253.6 899.7 7.0 1422.0 760.0 341.4 1398.6 9.0 1923.8 899.5 349.4 1847.612.0 2025.0 1038.5 436.4 2089.7 14.0 1213.4 516.0 16.0 1256.4 521.8 19.01741.5 560.9 21.0 1878.8 627.7 23.0 2057.0 690.9 26.0 953.4 28.0 847.130.0 1067.5 33.0 1200.6 35.0 1257.7 37.0 1497.7 41.0 2014.2

Example 29d. In Vivo Studies to Determine the Effect of 4-S and 12-S inHuman Ovarian Tumor Xenografts

The aim of this study was to compare the antitumoral activity of 4-S and12-S with the antitumoral activity of compound C by using a xenograftmodel of human ovarian cancer.

The tumor model used in this study was A2780.

Table 17 reports the volume evaluation of A2780 tumors in mice treatedwith placebo, compound C, 4-S, and 12-S. These results are also showedin FIG. 6.

TABLE 17 Median Tumor Volume (mm³) Days Control Compound C 4-S 12-S 0.0169.5 169.6 168.3 168.5 2.0 317.5 206.3 150.6 262.1 5.0 758.9 372.7175.9 628.6 7.0 1351.9 607.6 317.7 976.3 9.0 1675.8 696.2 281.9 1387.512.0 2025.0 855.6 372.1 1666.0 14.0 1293.9 709.2 2025.0 16.0 1683.5870.9 19.0 2137.5 1235.4 21.0 1453.3 23.0 1666.0 26.0 2025.0

Example 29e. In Vivo Studies to Determine the Effect of 4-S and 12-S inHuman Gastric Tumor Xenografts

The aim of this study was to compare the antitumoral activity of 4-S and12-S with the antitumoral activity of Compound C by using a xenograftmodel of human gastric cancer.

The tumor model used in this study was HGC27.

Table 18 reports tumor volume growth of HGC27 tumors in mice treatedwith placebo, compound C, 4-S, and 12-S. These results are also showedin FIG. 7.

TABLE 18 Median Tumor Volume (mm³) Days Control Compound C 4-S 12-S 0.0200.7 195.0 194.8 196.6 2.0 429.0 391.0 358.6 411.9 5.0 835.5 578.6515.3 834.1 7.0 1256.5 708.2 589.2 1176.6 9.0 1602.2 937.7 779.4 1531.612.0 2040.7 1169.5 980.8 2030.2 14.0 1496.8 1153.3 16.0 1690.6 1346.219.0 2004.0 1643.4 21.0 2004.7

Example 30. In Vivo Studies to Determine the Effect of 4-R in SeveralXenograft Models

4-R was provided in the form of freeze dried vials. 4-R cake wasreconstituted with water for infusion to a concentration of 0.5 mg/mL.The 4-R stock solution was further diluted in 5% dextrose solution forinjection to the dosing formulation concentration. The 4-R administereddose was 0.30 mg/kg.

Compound D was provided in the form of drug substance vials. Each vialwas reconstituted first by total dissolution in DMSO and then addingKolliphor ELP (BASF)/ethanol absolute (1:1, v/v) to a concentration of0.8 mg/mL. Further dilutions were made with a lactate buffer solution(pH=4.0) to the dosing formulation concentration. The Compound Dadministered dose was 0.5 mg/kg.

PM01183 was provided in the form of vials of lyophilized product. Eachvial was reconstituted with water for infusion to a concentration of 0.2mg/mL. Further dilutions were made with 5% glucose or 0.9% sodiumchloride solution for injection to the dosing formulationconcentrations. The administered dose was 0.18 mg/kg.

Placebo was provided in the form of lyophilised cake containing 100 mgSucrose+Potassium dihydrogen phosphate 6.8 mg+Phosphoric acid q.s. pH3.8-4.5 which was reconstituted with water for infusion.

In these experiments, 4-R, Compound D and PM01183, as well as placebo,were intravenously administered once per week for 3 consecutive weeks,on Days 0, 7 and 14, whenever it was possible.

Example 30a. In Vivo Studies to Determine the Effect of 4-R in HumanFibrosarcoma Xenografts

The aim of this study was to compare the antitumoral activity of 4-R andCompound D with the antitumoral activity of PM01183 by using a xenograftmodel of human sarcoma.

The tumor model used in this study was HT1080 cell line.

Table 19 reports the total diameter (tumor+leg) evaluation of HT1080tumors in mice treated with placebo, PM01183 and 4-R. These results arealso showed in FIG. 8.

TABLE 19 Total diameter (tumor + leg) (mm) Days Control PM01183 4-R 08.1 8.1 8.1 2 11.2 9.7 8.6 7 13.6 11.2 8.7 9 15.2 12.3 9.0 14 16.9 14.69.3 18 18.1 15.6 10.3 21 15.1 11.5 23 16.3 13.3 25 18.0 15.8 28 18.0

Table 20 reports the total diameter (tumor+leg) evaluation of HT1080tumors in mice treated with placebo, PM01183 and Compound D. Theseresults are also showed in FIG. 9.

TABLE 20 Total diameter (tumor + leg) (mm) Days Control PM01183 CompoundD 0 7.8 7.7 7.7 2 11.0 9.2 9.5 5 14.0 9.8 8.8 7 15.0 12.2 8.7 9 18.012.6 9.4 12 13.1 9.4 14 14.6 10.1 16 14.5 10.9 19 15.0 11.2 21 18.0 12.123 13.0 26 15.0 28 18.0

Example 30b. In Vivo Studies to Determine the Effect of 4-R in HumanBreast Xenografts

The aim of this study was to compare the antitumoral activity of 4-R andCompound D with the antitumoral activity of PM01183 by using a xenograftmodel of human breast cancer.

The tumor model used in this study was MDA-MB-231 cell line.

Table 21 reports the median tumor volume evaluation of MDA-MB-231 tumorsin mice treated with placebo, PM01183 and 4-R. These results are alsoshowed in FIG. 10.

TABLE 21 Median Tumor Volume (mm³) Days Control PM01183 4-R 0 130.6129.3 129.3 7 230.7 189.0 151.9 14 422.2 230.1 164.1 21 687.7 305.9136.8 28 1114.9 535.8 195.9 35 1555.3 819.7 294.2 42 2138.5 962.7 494.449 1301.3 843.8 52 2199.4 1042.5

Table 22 reports the volume evaluation of MDA-MB-231 tumors in micetreated with placebo, PM01183 and Compound D. These results are alsoshowed in FIG. 11.

TABLE 22 Median Tumor Volume (mm³) Days Control PM01183 Compound D 0129.2 129.6 129.5 7 284.0 185.9 147.9 14 564.3 290.8 186.4 21 686.0337.9 136.5 28 1068.6 507.4 290.7 35 1359.4 796.1 431.7 42 1533.7 1062.5770.1 49 1653.1 1416.3 970.0 56 2029.3 1673.3 1461.9 63 2060.8 1811.91526.4

Example 30c. In Vivo Studies to Determine the Effect of 4-R in HumanLung Tumor Xenografts

The aim of this study was to compare the antitumoral activity of 4-R andCompound D with the antitumoral activity of PM01183 by using a xenograftmodel of human lung cancer.

The tumor model used in this study was H-460 cell line.

Table 23 reports the volume evaluation of H460 tumors in mice treatedwith placebo, PM01183 and 4-R. These results are also showed in FIG. 12.

TABLE 23 Median Tumor Volume (mm³) Days Control PM01183 4-R 0 156.2156.7 155.5 2 290.9 227.3 223.3 7 1323.8 940.4 737.8 9 1816.9 1210.3861.0 11 2120.9 1433.8 1102.9 14 1529.5 1638.0 16 2028.6

Table 24 reports the volume evaluation of H460 tumors in mice treatedwith placebo, PM01183 and Compound D. These results are also showed inFIG. 13.

TABLE 24 Median Tumor Volume (mm³) Days Control PM01183 Compound D 0205.2 204.5 203.4 2 508.0 418.1 367.3 7 1355.8 1004.0 792.0 9 1682.11211.3 854.6 12 1938.6 1515.4 1026.7 14 2275.9 1633.3 1175.8 16 1723.91322.1 19 2112.3 1581.1 21 2409.4 1789.3 23 1966.5 26 2080.7

Example 30d. In Vivo Studies to Determine the Effect of 4-R in HumanOvarian Tumor Xenografts

The aim of this study was to compare the antitumoral activity of 4-R andCompound D with the antitumoral activity of PM01183 by using a xenograftmodel of human ovarian cancer.

The tumor model used in this study was A2780.

Table 25 reports the volume evaluation of A2780 tumors in mice treatedwith placebo, PM01183 and 4-R. These results are also showed in FIG. 14.

TABLE 25 Median Tumor Volume (mm³) Days Control PM01183 4-R 0 172.8175.5 175.2 5 896.6 671.2 611.4 7 1415.3 1048.9 1036.5 12 2205.3 2020.31992.0 14 2165.3

Table 26 reports the volume evaluation of A2780 tumors in mice treatedwith placebo, PM01183 and Compound D. These results are also showed inFIG. 15.

TABLE 26 Median Tumor Volume (mm³) Days Control PM01183 Compound D 0189.4 191.2 190.1 3 588.5 454.5 319.6 5 1086.0 772.1 514.4 7 1428.61161.5 897.4 10 2077.1 1615.6 1239.8 12 2163.1 1703.0 1656.2 14 2029.31951.7 17 2121.7 19 2068.6

Example 30e. In Vivo Studies to Determine the Effect of 4-R in HumanGastric Tumor Xenografts

The aim of this study was to compare the antitumoral activity of 4-R andCompound D with the antitumoral activity of PM01183 by using a xenograftmodel of human gastric cancer.

The tumor model used in this study was HGC27.

Table 27 reports tumor volume growth of HGC27 tumors in mice treatedwith placebo, PM01183 and 4-R. These results are also showed in FIG. 16.

TABLE 27 Median Tumor Volume (mm³) Days Control PM01183 4-R 0 174.6171.6 173.0 2 319.1 317.5 266.8 5 632.5 404.0 370.7 7 1046.0 485.7 418.59 1359.1 604.6 627.8 12 1863.8 760.8 713.5 14 2115.0 789.6 837.0 16719.5 867.1 19 895.9 1040.2 21 1051.3 1229.8 26 1901.2 1784.5 28 2028.92073.6

Table 28 reports tumor volume growth of HGC27 tumors in mice treatedwith placebo, PM01183 and Compound D. These results are also showed inFIG. 17.

TABLE 28 Median Tumor Volume (mm³) Days Control PM01183 Compound D 0142.3 169.5 157.4 2 286.5 372.4 327.6 5 527.7 474.1 439.6 7 821.4 571.8418.7 9 1130.9 787.9 567.9 12 1547.8 951.1 537.0 14 1868.5 1064.4 654.616 1887.0 1346.1 672.4 19 2162.3 1691.8 843.0 21 1920.0 842.7 23 2011.4963.7 26 2102.2 1203.3 28 1589.7 30 1777.6 33 2146.2

Example 31. In Vivo Studies to Determine the Effect of 12-R in SeveralXenograft Models

12-R was provided in the form of freeze dries vials. 12-R cake wasreconstituted with water for infusion to a concentration of 0.5 mg/mL.The 12-R stock solution was further diluted in 5% dextrose solution forinjection to the dosing formulation concentration. The 12-R administereddose was 0.05 mg/kg.

Compound D was provided in the form of drug substance vials. Each vialwas reconstituted first by total dissolution in DMSO and then addingKolliphor ELP (BASF)/ethanol absolute (1:1, v/v) to a concentration of0.8 mg/mL. Further dilutions were made with a lactate buffer solution(pH=4.0) to the dosing formulation concentration. The Compound Dadministered dose was 0.5 mg/kg.

Placebo was provided in the form of lyophilised cake containing 100 mgSucrose+Potassium dihydrogen phosphate 6.8 mg+Phosphoric acid q.s. pH3.8-4.5 which was reconstituted with water for infusion.

In these experiments, 12-R, Compound D, as well as placebo, wereintravenously administered once per week for 3 consecutive weeks, onDays 0, 7 and 14, whenever it was possible.

Example 31a. In Vivo Studies to Determine the Effect of 12-R in HumanFibrosarcoma Xenografts

The aim of this study was to compare the antitumoral activity of 12-Rwith the antitumoral activity of Compound D by using a xenograft modelof human sarcoma.

The tumor model used in this study was HT1080 cell line.

Table 29 reports the total diameter (tumor+leg) evaluation of HT1080tumors in mice treated with placebo, Compound D and 12-R. These resultsare also showed in FIG. 18.

TABLE 29 Total diameter (tumor + leg) (mm) Days Control Compound D 12-R0.0 7.5 7.5 7.5 2.0 9.4 8.2 8.9 5.0 11.4 7.5 8.8 7.0 12.1 7.4 9.5 9.013.2 8.1 9.5 12.0 14.5 7.9 11.0 14.0 15.2 7.7 11.7 16.0 15.9 8.8 12.919.0 18.0 10.2 13.5 21.0 11.2 15.5 23.0 12.2 18.0 27.0 13.2 30.0 14.633.0 16.3 35.0 18.0

Example 31b. In Vivo Studies to Determine the Effect of 12-R in HumanBreast Xenografts

The aim of this study was to compare the antitumoral activity of 12-Rwith the antitumoral activity of Compound D by using a xenograft modelof human breast cancer.

The tumor model used in this study was MDA-MB-231 cell line.

Table 30 reports the median tumor volume evaluation of MDA-MB-231 tumorsin mice treated with placebo, Compound D and 12-R. These results arealso showed in FIG. 19.

TABLE 30 Median Tumor Volume (mm³) Days Control Compound D 12-R 0.0149.4 149.6 149.8 2.0 240.0 217.2 223.0 5.0 325.1 284.5 296.1 7.0 407.8310.0 378.3 9.0 514.8 325.5 472.7 12.0 648.1 268.4 609.9 14.0 799.0237.7 782.5 16.0 1002.5 261.2 972.4 19.0 1233.9 251.3 1211.0 21.0 1539.1219.9 1463.4 23.0 2006.5 221.8 1756.5 26.0 2027.7 245.5 2028.6 28.0320.3 30.0 401.6 33.0 545.8 35.0 629.2 37.0 670.7 40.0 669.9 42.0 696.344.0 798.1 47.0 857.7

Example 31c. In Vivo Studies to Determine the Effect of 12-R in HumanLung Tumor Xenografts

The aim of this study was to compare the antitumoral activity of 12-Rwith the antitumoral activity of Compound D by using three differentxenograft models of human lung cancer. These models correspond tonon-small cell lung cancer (H460 cell line and to small cell lung cancer(H526 and H82 cell lines).

Table 31 reports the volume evaluation of H460 tumors in mice treatedwith placebo, Compound D and 12-R. These results are also showed in FIG.20.

TABLE 31 Median Tumor Volume (mm³) Days Control Compound D 12-R 0.0187.4 187.2 187.0 2.0 577.5 329.7 410.7 5.0 1352.0 559.4 796.7 7.01642.9 756.5 1167.9 9.0 2025.0 971.9 1360.3 12.0 1370.9 1666.0 14.01626.8 2025.0 16.0 2025.0

Table 32 reports the median tumor volume evaluation of H526 tumors inmice treated with placebo, compound D and 12-R. The results are alsoshown in FIG. 21.

TABLE 32 Median Tumor Volume (mm³) Days Control Compound D 12-R 0.0217.20 216.1 214.20 2.0 410.70 240.9 404.50 4.0 778.50 99.3 680.50 7.01083.20 56.7 995.20 9.0 1371.00 62.5 1290.50 11.0 1782.00 62.5 1568.0014.0 2025.00 32.0 2025.00 16.0 4.0 21.0 4.0 28.0 4.0 35.0 4.0 42.0 4.049.0 4.0

Table 33 reports the median tumor volume evaluation of H82 tumors inmice treated with placebo, compound D and 12-R. The results are alsoshown in FIG. 22.

TABLE 33 Median Tumor Volume (mm³) Days Control Compound D 12-R 0.0171.60 169.4 170.50 2.0 439.40 340.6 381.40 5.0 1024.70 443.3 793.20 7.01422.00 496.2 1187.20 9.0 1923.80 614.1 1699.30 12.0 2025.00 665.52125.60 14.0 1041.6 16.0 1151.2 19.0 1516.7 21.0 1748.0

Example 31d. In Vivo Studies to Determine the Effect of 12-R in HumanOvarian Tumor Xenografts

The aim of this study was to compare the antitumoral activity of 12-Rwith the antitumoral activity of Compound D by using a xenograft modelof human ovarian cancer.

The tumor model used in this study was A2780.

Table 34 reports the volume evaluation of A2780 tumors in mice treatedwith placebo, Compound D and 12-R. These results are also showed in FIG.23.

TABLE 34 Median Tumor Volume (mm³) Days Control Compound D 12-R 0.0169.5 168.8 169.6 2.0 317.5 225.7 302.8 5.0 758.9 256.6 786.5 7.0 1351.9473.8 1113.3 9.0 1675.8 633.6 1490.6 12.0 2025.0 822.8 2025.00 14.01129.3 2025.00 16.0 1198.6 19.0 1649.6 21.0 2025.0

Example 31e. In Vivo Studies to Determine the Effect of 12-R in HumanGastric Tumor Xenografts

The aim of this study was to compare the antitumoral activity of 12-Rwith the antitumoral activity of Compound D by using a xenograft modelof human gastric cancer.

The tumor model used in this study was HGC27.

Table 35 reports tumor volume growth of HGC27 tumors in mice treatedwith placebo, Compound D and 12-R. These results are also showed in FIG.24.

TABLE 35 Median Tumor Volume (mm³) Days Control Compound D 12-R 0.0200.7 194.0 193.3 2.0 429.0 324.2 413.3 5.0 835.5 561.6 809.1 7.0 1256.5504.2 1261.5 9.0 1602.2 584.2 1589.5 12.0 2040.7 767.7 2017.9 14.01056.8 2034.9 16.0 1440.2 19.0 1717.9 21.0 2043.4

Example 32. In Vivo Studies to Determine the Effect of 19-S in SeveralXenograft Models

19-S was provided in the form of freeze dried vials. 19-S cake wasreconstituted with water for infusion to a concentration of 0.5 mg/mL.The 19-S stock solution was further diluted in 5% dextrose solution forinjection to the dosing formulation concentration. The 19-S administereddose was 0.75 mg/kg.

PM01183 was provided in the form of vials of lyophilized product. Eachvial was reconstituted with water for infusion to a concentration of 0.2mg/mL. The PM01183 stock solution was further diluted in 5% glucosesolution for injection to the dosing formulation concentrations. Theadministered dose was 0.18 mg/kg.

Placebo was provided in the form of lyophilised cake containing 100 mgSucrose+Potassium dihydrogen phosphate 6.8 mg+Phosphoric acid q.s. pH3.8-4.5 which was reconstituted with water for infusion.

In these experiments, 19-S and PM01183, as well as placebo, wereintravenously administered once per week for 3 consecutive weeks, onDays 0, 7 and 14, whenever it was possible.

Example 32a. In Vivo Studies to Determine the Effect of 19-S in HumanFibrosarcoma Xenografts

The aim of this study was to compare the antitumoral activities of 19-Sand PM01183 by using a xenograft model of human sarcoma.

The tumor model used in this study was HT1080 cell line.

Table 36 reports the total diameter (tumor+leg) evaluation of HT1080tumors in mice treated with placebo, PM01183 and 19-S. These results arealso showed in FIG. 25.

TABLE 36 Total diameter (tumor + leg) (mm) Days Control PM01183 19-S 08.4 8.4 8.2 2 10.9 9.8 8.4 5 14.8 9.7 7.8 7 15.9 11.4 9.5 9 18.0 12.79.9 12 13.7 10.7 14 14.6 11.3 16 15.5 11.9 19 15.6 13.4 21 18.0 14.4 2318.0

Example 32b. In Vivo Studies to Determine the Effect of 19-S in HumanBreast Adenocarcinoma Xenografts

The aim of this study was to compare the antitumoral activities of 19-Sand PM01183 by using a xenograft model of human breast cancer.

The tumor model used in this study was MDA-MB-231 cell line.

Table 37 reports the median tumor volume evaluation of MDA-MB-231 tumorsin mice treated with placebo, PM01183 and 19-S. These results are alsoshowed in FIG. 26.

TABLE 37 Median Tumor Volume (mm³) Days Control PM01183 19-S 0 132.6134.3 133.6 4 194.1 177.2 157.2 7 248.2 186.3 142.6 11 377.6 250.7 133.914 461.3 266.1 117.3 18 679.2 327.7 79.3 21 753.2 391.0 89.2 25 909.2493.1 120.6 28 1090.7 627.3 144.4 32 1433.4 789.0 246.1 36 1887.5 1022.0419.3 39 1785.2 1294.2 593.7 42 2081.5 1643.3 945.9 46 2137.5 1658.9985.3 49 1938.0 1211.5 53 1324.3 56 1703.9 60 1793.3 63 1603.0 70 2324.2

Example 32c. In Vivo Studies to Determine the Effect of 19-S in HumanLung Cancer Xenografts

The aim of this study was to compare the antitumoral activities of 19-Sand PM01183 by using a xenograft model of human lung cancer.

The tumor model used in this study was H-460 cell line.

Table 38 reports the median tumor volume evaluation of H-460 tumors inmice treated with placebo, PM01183 and 19-S. These results are alsoshowed in FIG. 27.

TABLE 38 Median Tumor Volume (mm³) Days Control PM01183 19-S 0 197.0196.3 196.9 2 529.5 457.0 364.0 4 1057.4 861.5 624.9 7 1582.5 1280.2966.5 9 2094.8 1424.9 1078.2 11 1969.9 1449.0 14 1761.5

Example 32d. In Vivo Studies to Determine the Effect of 19-S in HumanOvarian Tumor Xenografts

The aim of this study was to compare the antitumoral activities of 19-Sand PM01183 by using a xenograft model of human ovarian cancer.

The tumor model used in this study was A2780.

Table 39 reports the median tumor volume evaluation of A2780 tumors inmice treated with placebo, PM01183 and 19-S. These results are alsoshowed in FIG. 28.

TABLE 39 Median Tumor Volume (mm³) Days Control PM01183 19-S 0 163.4163.6 164.4 2 287.1 235.5 187.9 4 568.7 463.2 205.4 7 1211.3 986.3 513.69 1633.7 1451.4 650.6 11 2047.8 2062 659.8 14 1236.2 18 1575.9 23 1895.725 2177.0

Example 32e. In Vivo Studies to Determine the Effect of 19-S in HumanGastric Tumor Xenografts

The aim of this study was to compare the antitumoral activities of 19-Sand PM01183 by using a xenograft model of human gastric cancer.

The tumor model used in this study was HGC27.

Table 40 reports the median tumor volume evaluation of HGC27 tumors inmice treated with placebo, PM01183 and 19-S. These results are alsoshowed in FIG. 29.

TABLE 40 Median Tumor Volume (mm³) Days Control PM01183 19-S 0 178.3177.6 181.5 2 409 395.6 404.6 5 907.4 572.4 600.3 7 1283.6 766.6 660.3 91664 950.7 787.5 14 2102.8 1199.4 864.4 16 1353.1 882.4 19 1294.3 925.221 1335.1 893.6 23 1320.3 874.4 26 1364.5 932.1 30 1671.9 1547.8 332009.2 2020.4

Example 33. In Vivo Studies to Determine the Effect of 19-R in SeveralXenograft Models

19-R was provided in the form of freeze dried vials. 19-R cake wasreconstituted with water for infusion to a concentration of 0.5 mg/mL.The 19-R stock solution was further diluted in 5% dextrose solution forinjection to the dosing formulation concentration. The 19-R administereddose was 0.15 mg/kg.

PM01183 was provided in the form of vials of lyophilized product. Eachvial was reconstituted with water for infusion to a concentration of 0.2mg/mL. The PM01183 stock solution was further diluted in 5% glucosesolution for injection to the dosing formulation concentrations. Theadministered dose was 0.18 mg/kg.

Placebo was provided in the form of lyophilised cake containing 100 mgSucrose+Potassium dihydrogen phosphate 6.8 mg+Phosphoric acid q.s. pH3.8-4.5 which was reconstituted with water for infusion.

In these experiments, 19-R and PM01183, as well as placebo, wereintravenously administered once per week for 3 consecutive weeks, onDays 0, 7 and 14, whenever it was possible.

Example 33a. In Vivo Studies to Determine the Effect of 19-R in HumanFibrosarcoma Xenografts

The aim of this study was to compare the antitumoral activity of 19-Rwith the antitumoral activity of PM01183 by using a xenograft model ofhuman sarcoma.

The tumor model used in this study was HT1080 cell line.

Table 41 reports the total diameter (tumor+leg) evaluation of HT-1080tumors in mice treated with placebo, PM01183 and 19-R. These results arealso showed in FIG. 30.

TABLE 41 Total diameter (tumor + leg) (mm) Days Control PM01183 19-R 08.4 8.4 8.3 2 10.9 9.8 9.4 5 14.8 9.7 8.0 7 15.9 11.4 7.2 9 18.0 12.77.8 12 13.7 7.8 14 14.6 8.4 16 15.5 8.2 19 15.6 11.3 21 18.0 12.2 2313.3 26 15.2 28 18.0

Example 33b. In Vivo Studies to Determine the Effect of 19-R in HumanBreast Adenocarcinoma Xenografts

The aim of this study was to compare the antitumoral activity of 19-Rwith the antitumoral activity of PM01183 by using a xenograft model ofhuman breast cancer.

The tumor model used in this study was MDA-MB-231 cell line.

Table 42 reports the median tumor volume evaluation of MDA-MB-231 tumorsin mice treated with placebo, PM01183 and 19-R. These results are alsoshowed in FIG. 31.

TABLE 42 Median Tumor Volume (mm³) Days Control PM01183 19-R 0 132.6134.3 132.5 4 194.1 177.2 189.3 7 248.2 186.3 151.9 11 377.6 250.7 167.514 461.3 266.1 152.6 18 679.2 327.7 162.2 21 753.2 391.0 201.2 25 909.2493.1 208.5 28 1090.7 627.3 274.8 32 1433.4 789.0 355.8 36 1887.5 1022.0513.8 39 1785.2 1294.2 793.7 42 2081.5 1643.3 1012.2 46 2137.5 1658.91188.5 49 1938.0 1380.7 53 1568.0 56 1862.6 60 2129.4

Example 33c. In Vivo Studies to Determine the Effect of 19-R in HumanLung Tumor Xenografts

The aim of this study was to compare the antitumoral activity of 19-Rwith the antitumoral activity of PM01183 by using a xenograft model ofhuman lung cancer.

The tumor model used in this study was H-460 cell line.

Table 43 reports the median tumor volume evaluation of H460 tumors inmice treated with placebo, PM01183 and 19-R. These results are alsoshowed in FIG. 32.

TABLE 43 Median Tumor Volume (mm³) Days Control PM01183 19-R 0 197.0196.3 196.8 2 529.5 457.0 418.7 4 1057.4 861.5 697.2 7 1582.5 1280.2911.7 9 2094.8 1424.9 1111.5 11 1969.9 1281.3 14 1478.7 16 1594.0

Example 33d. In Vivo Studies to Determine the Effect of 19-R in HumanOvarian Tumor Xenografts

The aim of this study was to compare the antitumoral activity of 19-Rwith the antitumoral activity of PM01183 by using a xenograft model ofhuman ovarian cancer.

The tumor model used in this study was A2780.

Table 44 reports the median tumor volume evaluation of A2780 tumors inmice treated with placebo, PM01183 and 19-R. These results are alsoshowed in FIG. 33.

TABLE 44 Median Tumor Volume (mm³) Days Control PM01183 19-R 0 163.4163.6 162.8 2 287.1 236.5 212.9 4 568.7 463.2 368.5 7 1211.3 986.3 841.39 1633.7 1451.4 1138.9 11 2047.8 2062.0 1519.9 14 2056.0

Example 33e. In Vivo Studies to Determine the Effect of 19-R in HumanGastric Tumor Xenografts

The aim of this study was to compare the antitumoral activity of 19-Rwith the antitumoral activity of PM01183 by using a xenograft model ofhuman gastric cancer.

The tumor model used in this study was HGC27.

Table 45 reports the median tumor volume evaluation of HGC-27 tumors inmice treated with placebo, PM01183 and 19-R. These results are alsoshowed in FIG. 34.

TABLE 45 Median Tumor Volume (mm³) Days Control PM01183 19-R 0 178.3177.6 182.0 2 409.0 395.6 414.9 5 907.4 572.4 735.0 7 1283.6 766.6 901.29 1664.0 950.7 1048.1 14 2102.8 1199.4 1293.9 16 1353.1 1488.8 19 1294.31668.3 21 1335.1 1845.0 23 1320.3 2025.0 26 1364.5 30 1671.9 33 2009.2

Example 34. In Vivo Studies to Determine the Effect of 39-S in SeveralXenograft Models

Compound 39-S and C were provided in the form of freeze-dried vials oflyophilized product. Each vial was reconstituted with sterile water forinjection to a concentration of 0.5 mg/mL. Further dilutions were madewith 5% dextrose solution for injection to the dosing formulationconcentration. The administered doses of 39-S and C were 1.25 and 3mg/Kg, respectively.

Placebo was provided in the forms of vials of lyophilised product. Eachvial (sucrose 200 mg+potassium dihydrogen phosphate 13.6 mg+phosphoricacid q.s. pH 3.8-4.5) was reconstituted with sterile water for injection(2 mL). Further dilutions were made with 5% dextrose solution forinjection.

In these experiments, 39-S and compound C, as well as placebo, wereintravenously administered on a weekly schedule at a volume of 10 mL/Kg.

Example 34a. In Vivo Studies to Determine the Effect of 39-S in HumanFibrosarcoma Xenografts

The aim of this study was to evaluate the antitumoral activity ofcompound 39-S by comparison with the antitumoral activity of compound Cby using a xenograft model of human sarcoma.

The tumor model used in this study was HT1080 cell line.

Table 46 reports the total diameter (tumor+leg) evaluation of HT1080tumors in mice treated with placebo, compound C and 39-S. These resultsare also showed in FIG. 35.

TABLE 46 Total diameter (tumor + leg) (mm) Days Control 39-S Compound C0 7.5 7.5 7.5 2 9.4 7.9 8.8 5 11.4 6.4 9.0 7 12.1 6.8 9.6 9 13.2 6.910.2 12 14.5 6.6 10.2 14 15.2 6.4 11.2 16 15.9 6.8 12.4 19 18.0 7.0 13.321 7.0 15.2 23 8.5 18.0 27 10.8 30 12.5 33 14.3 35 15.3 37 18.0

Example 34b. In Vivo Studies to Determine the Effect of 39-S in HumanBreast Adenocarcinoma Xenografts

The aim of this study was to compare the antitumoral activities of 39-Sand compound C by using a xenograft model of human breast cancer.

The tumor model used in this study was MDA-MB-231 cell line.

Table 47 reports the median tumor volume evaluation of MDA-MB-231 tumorsin mice treated with placebo, compound C and 39-S. These results arealso showed in FIG. 36.

TABLE 47 Median Tumor Volume (mm³) Days Control 39-S Compound C 0 149.4151.0 149.4 2 240.0 209.3 217.1 5 325.1 290.9 281.3 7 407.8 301.8 338.69 514.8 300.8 385.1 12 648.1 278.7 400.4 14 799.0 249.7 436.9 16 1002.5243.6 585.7 19 1233.9 248.3 774.7 21 1539.1 250.0 965.9 23 2006.5 260.31215.2 26 2027.7 304.9 1503.2 28 337.1 1785.3 30 451.3 2037.1 33 584.135 683.4 37 784.7 40 937.4 42 1060.5 44 1170.5 47 1112.9 49 1138.6 511283.2 54 1415.1 56 1518.7 58 1728.5 61 2017.9

Example 34c. In Vivo Studies to Determine the Effect of 39-S in HumanLung Cancer Xenografts

The aim of this study was to compare the antitumoral activity of 39-Swith the antitumoral activity of compound C by using three differentxenograft models of human lung cancer. These models correspond tonon-small cell lung cancer (H-460 cell line) and to small cell lungcancer (H526 and H82 cell lines).

Table 48 reports the median tumor volume evaluation of H460 tumors inmice treated with placebo, compound C and 39-S. These results are alsoshowed in FIG. 37.

TABLE 48 Median Tumor Volume (mm³) Days Control 39-S Compound C 0 187.4187.8 186.1 2 577.5 314.4 395.4 5 1352.0 584.1 665.9 7 1642.9 831.2929.5 9 2025.0 841.0 1063.7 12 1008.0 1436.5 14 1309.8 2025.0 16 1470.02025.0 19 2025.0

Table 49 reports the median tumor volume evaluation of H526 tumors inmice treated with placebo, compound C and 39-S. These results are alsoshowed in FIG. 38.

TABLE 49 Median Tumor Volume (mm³) Days Control 39-S Compound C 0 217.2214.5 217.9 2 410.7 260.3 262.4 4 778.5 80.0 108.3 7 1083.2 46.2 129.8 91371.0 32.0 85.9 11 1782.0 32.0 52.3 14 2025.0 4.0 54.1 16 4.0 47.3 214.0 4.0 28 4.0 4.0 35 4.0 4.0 42 4.0 62.5 49 4.0 53.5 56 4.0 70.0 63 4.0132.3 70 4.0 368.5 77 4.0 465.8 84 4.0 107.4 91 4.0 130.0 98 4.0 4.0 1054.0 4.0 112 4.0 4.0 119 4.0 4.0 126 4.0 4.0 133 4.0 4.0 140 4.0 4.0 1474.0 4.0 165 4.0 4.0 175 4.0 4.0 191 4.0 4.0 205 4.0 4.0

Table 50 reports the median tumor volume evaluation of H82 tumors inmice treated with placebo, compound C and 39-S. These results are alsoshowed in FIG. 39.

TABLE 50 Median Tumor Volume (mm³) Days Control 39-S Compound C 0 171.6170.3 170.5 2 439.4 325.2 265.3 5 1024.7 430.8 488.7 7 1422.0 466.2760.0 9 1923.8 544.3 899.5 12 2025.0 640.3 1038.5 14 711.2 1213.4 16802.7 1256.4 19 916.0 1741.5 21 1047.2 1878.8 23 1189.1 2057.0 26 1497.228 1741.8 30 1731.7 33 2029.4

Example 34d. In Vivo Studies to Determine the Effect of 39-S in HumanOvarian Tumor Xenografts

The aim of this study was to compare the antitumoral activity of 39-Swith the antitumoral activity of compound C by using a xenograft modelof human ovarian cancer.

The tumor model used in this study was A2780.

Table 51 reports the volume evaluation of A2780 tumors in mice treatedwith placebo, compound C and 39-S. These results are also showed in FIG.40.

TABLE 51 Median Tumor Volume (mm³) Day Control 39-S Compound C 0 169.5170.5 169.6 2 317.5 206.5 206.3 5 758.9 163.4 372.7 7 1351.9 298.6 607.69 1675.8 317.4 696.2 12 2025.0 378.2 855.6 14 668.5 1293.9 16 853.51683.5 19 1415.5 2137.5 21 1519.2 23 1666.0 30 2025.0

Example 34e. In Vivo Studies to Determine the Effect of 39-S in HumanGastric Tumor Xenografts

The aim of this study was to compare the antitumoral activity of 39-Swith the antitumoral activity of compound C by using a xenograft modelof human gastric cancer.

The tumor model used in this study was HGC27.

Table 52 reports tumor volume growth of HGC27 tumors in mice treatedwith placebo, compound C, and 39-S. These results are also showed inFIG. 41.

TABLE 52 Median Tumor Volume (mm³) Days Control 39-S Compound C 0 200.7195.6 195.0 2 429.0 356.3 391.0 5 835.5 469.7 578.6 7 1256.5 467.8 708.29 1602.2 575.2 937.7 12 2040.7 611.1 1169.5 14 637.3 1496.8 16 690.41690.6 19 701.8 2004.0 21 697.4 1741.4 23 715.5 2056.4 26 898.1 281163.4 30 1409.3 33 1450.5 35 1708.5 37 1804.4 40 2075.2

Example 35. In Vivo Studies to Determine the Effect of 47-R in SeveralXenograft Models

Compound 47-R was provided in the form of freeze-dried vials oflyophilized product. Each vial was reconstituted with sterile water forinjection to a concentration of 0.5 mg/mL. Further dilutions were madewith 5% dextrose solution for injection to the dosing formulationconcentration. 47-R administered dose was 0.1 mg/Kg.

Compound D was provided in the form of powder drug substance. Each vialwas reconstituted first by total dissolution in DMSO (Fisher) and thenadding Kolliphor ELP (Basf)/ethanol absolute (Merk) (1:1, v/v) to aconcentration of 0.8 mg/mL. Further dilutions were made with a lactatebuffer solution (pH=4.0) to the dosing formulation concentration.Compound D administered dose was 0.5 mg/Kg.

Placebo was provided in the form of vials of lyophilised product. Eachvial (sucrose 200 mg+potassium dihydrogen phosphate 13.6 mg+phosphoricacid q.s. pH 3.8-4.5) was reconstituted with sterile water for injection(2 mL). Further dilutions were made with 5% dextrose solution forinjection.

In these experiments, 47-R and compound D, as well as placebo, wereintravenously administered on a weekly schedule at a volume of 10 mL/Kg.

Example 35a. In Vivo Studies to Determine the Effect of 47-R in HumanFibrosarcoma Xenografts

The aim of this study was to evaluate the antitumoral activity ofcompound 47-R by comparison with the antitumoral activity of compound Dby using a xenograft model of human sarcoma.

The tumor model used in this study was HT1080 cell line.

Table 53 reports the total diameter (tumor+leg) evaluation of HT1080tumors in mice treated with placebo, compound D and 47-R. These resultsare also showed in FIG. 42.

TABLE 53 Total diameter (tumor + leg) (mm) Days Control 47-R Compound D0 7.5 7.5 7.5 2 9.4 8.9 8.2 5 11.4 10.1 7.5 7 12.1 10.5 7.4 9 13.2 11.58.1 12 14.5 13.5 7.9 14 15.2 13.9 7.7 16 15.9 14.6 8.8 19 18.0 18.0 10.221 11.2 23 12.2 27 13.2 30 14.6 33 16.3 35 18.0

Example 35b. In Vivo Studies to Determine the Effect of 47-R in HumanBreast Adenocarcinoma Xenografts

The aim of this study was to compare the antitumoral activities of 47-Rand compound D by using a xenograft model of human breast cancer.

The tumor model used in this study was MDA-MB-231 cell line.

Table 54 reports the median tumor volume evaluation of MDA-MB-231 tumorsin mice treated with placebo, compound D and 47-R. These results arealso showed in FIG. 43.

TABLE 54 Median Tumor Volume (mm³) Days Control 47-R Compound D 0 149.4150.5 149.6 2 240.0 225.3 217.2 5 325.1 323.2 284.5 7 407.8 405.0 310.09 514.8 495.9 325.5 12 648.1 594.1 268.4 14 799.0 769.5 237.7 16 1002.51009.5 261.2 19 1233.9 1298.0 251.3 21 1539.1 1580.7 219.9 23 2006.52006.5 221.8 26 2027.7 2032.1 245.5 28 320.3 30 401.6 33 545.8 35 629.237 670.7 40 669.9 42 696.3 44 798.1 47 857.7 49 870.7 51 925.8 54 1005.456 1064.2 58 1235.6 61 1367.8 63 1553.7 65 2017.9

Example 35c. In Vivo Studies to Determine the Effect of 47-R in HumanLung Cancer Xenografts

The aim of this study was to compare the antitumoral activity of 47-Rwith the antitumoral activity of compound D by using three differentxenograft models of human lung cancer. These models correspond tonon-small cell lung cancer (H-460 cell line) and to small cell lungcancer (H526 and H82 cell lines).

Table 55 reports the median tumor volume evaluation of H460 tumors inmice treated with placebo, compound D and 47-R. These results are alsoshowed in FIG. 44.

TABLE 55 Median Tumor Volume (mm³) Days Control 47-R Compound D 0 187.4185.8 187.2 2 577.5 508.1 329.7 5 1352.0 979.3 559.4 7 1642.9 1280.0756.5 9 2025.0 1543.1 971.9 12 1764.0 1370.9 14 1845.5 1626.8 16 2025.0

Table 56 reports the median tumor volume evaluation of H526 tumors inmice treated with placebo, compound D and 47-R. These results are alsoshowed in FIG. 45.

TABLE 56 Median Tumor Volume (mm³) Days Control 47-R Compound D 0 217.2211.5 216.1 2 410.7 367.9 240.9 4 778.5 583.7 99.3 7 1083.2 941.7 56.7 91371.0 1305.2 62.5 11 1782.0 1484.7 62.5 14 2025.0 2025.0 32.0 16 4.0 214.0 28 4.0 35 4.0 42 4.0 49 4.0 56 4.0 63 4.0 70 4.0 77 4.0 84 4.0 914.0 98 4.0 105 4.0 112 4.0 119 4.0 126 4.0 133 4.0 140 4.0 147 4.0 1654.0 175 4.0 191 4.0 205 4.0

Table 57 reports the median tumor volume evaluation of H82 tumors inmice treated with placebo, compound D and 47-R. These results are alsoshowed in FIG. 46.

TABLE 57 Median Tumor Volume (mm³) Days Control 47-R Compound D 0 171.6169.0 169.4 2 439.4 371.6 340.6 5 1024.7 888.8 443.3 7 1422.0 1314.2496.2 9 1923.8 1811.0 614.1 12 2025.0 2055.4 665.5 14 1041.6 16 1151.219 1516.7 21 1748.0

Example 35d. In Vivo Studies to Determine the Effect of 47-R in HumanOvarian Tumor Xenografts

The aim of this study was to compare the antitumoral activity of 47-Rwith the antitumoral activity of compound D by using a xenograft modelof human ovarian cancer.

The tumor model used in this study was A2780.

Table 58 reports the volume evaluation of A2780 tumors in mice treatedwith placebo, compound D and 47-R. These results are also showed in FIG.47.

TABLE 58 Median Tumor Volume (mm³) Days Control 47-R Compound D 0 169.5170.6 168.8 2 317.5 280.6 225.7 5 758.9 653.9 256.6 7 1351.9 848.7 473.89 1675.8 1569.1 633.6 12 2025.0 1764.0 822.8 14 1666.0 1129.3 16 2025.01198.6 19 1649.6 21 2025.0

Example 35e. In Vivo Studies to Determine the Effect of 47-R in HumanGastric Tumor Xenografts

The aim of this study was to compare the antitumoral activity of 47-Rwith the antitumoral activity of compound D by using a xenograft modelof human gastric cancer.

The tumor model used in this study was HGC27.

Table 59 reports tumor volume growth of HGC27 tumors in mice treatedwith placebo, compound D, and 47-R. These results are also showed inFIG. 48.

TABLE 59 Median Tumor Volume (mm³) Days Control 47-R Compound D 0 200.7194.0 194.0 2 429.0 359.4 324.2 5 835.5 774.8 561.6 7 1256.5 1155.4504.2 9 1602.2 1474.7 584.2 12 2040.7 1870.2 767.7 14 2031.3 1056.8 162075.2 1440.2 19 1717.9 21 2043.4

Example 36. In Vivo Studies to Determine the Effect of 32 in SeveralXenograft Models

Compounds 32 and ET-736 were provided in the form of freeze-dried vialsof lyophilized product. Each vial was reconstituted with sterile waterfor injection to a concentration of 0.5 mg/mL. Further dilutions weremade with 5% dextrose solution for injection to the dosing formulationconcentration. The administered dose of 32 and ET-736 was 0.5 mg/Kg.

Placebo was provided in the form of lyophilised product. Each vial(sucrose 200 mg+potassium dihydrogen phosphate 13.6 mg+phosphoric acidq.s. pH 3.8-4.5) was reconstituted with sterile water for injection (2mL). Further dilutions were made with 5% dextrose solution forinjection.

In these experiments, 32 and ET-736, as well as placebo, wereintravenously administered on a weekly schedule at a volume of 10 mL/Kg.

Example 36a. In Vivo Studies to Determine the Effect of 32 in HumanFibrosarcoma Xenografts

The aim of this study was to evaluate the antitumoral activity ofcompound 32 by comparison with the antitumoral activity of ET-736 byusing a xenograft model of human sarcoma.

The tumor model used in this study was HT-1080 cell line.

Table 60 reports the total diameter (tumor+leg) evaluation of HT1080tumors in mice treated with placebo, ET-736 and 32. These results arealso showed in FIG. 49.

TABLE 60 Total diameter (tumor + leg) (mm) Days Control 32 ET-736 0 7.57.5 7.4 2 9.4 8.9 8.3 5 11.4 8.2 7.1 7 12.1 8.8 7.6 9 13.2 10.0 7.4 1214.5 8.8 7.0 14 15.2 10.8 7.1 16 15.9 11.8 7.4 19 18.0 12.0 8.4 21 14.08.6 23 13.8 10.0 27 13.6 10.9 30 15.5 13.2 33 18.0 14.3 35 15.2 37 15.840 16.6 42 18.0

Example 36b. In Vivo Studies to Determine the Effect of 32 in HumanBreast Adenocarcinoma Xenografts

The aim of this study was to compare the antitumoral activities of 32and ET-736 by using a xenograft model of human breast cancer.

The tumor model used in this study was MDA-MB-231 cell line.

Table 61 reports the median tumor volume evaluation of MDA-MB-231 tumorsin mice treated with placebo, ET-736 and 32. These results are alsoshowed in FIG. 50.

TABLE 61 Median Tumor Volume (mm³) Days Control 32 ET-736 0 149.4 150.2150.0 2 240.0 233.6 237.7 5 325.1 310.6 302.1 7 407.8 386.1 364.9 9514.8 437.5 404.6 12 648.1 493.4 395.4 14 799.0 560.3 398.3 16 1002.5649.5 447.2 19 1233.9 853.0 485.0 21 1539.1 1017.5 536.3 23 2006.51263.2 669.8 26 2027.7 1487.7 778.9 28 1726.6 1046.1 30 1892.6 1315.9 332082.8 1664.9 35 2007.7

Example 36c. In Vivo Studies to Determine the Effect of 32 in Human LungCancer Xenografts

The aim of this study was to compare the antitumoral activities of 32and ET-736 by using three different xenograft models of human lungcancer. These models correspond to non-small cell lung cancer (H-460cell line) and to small cell lung cancer (H526 and H82 cell lines).

Table 62 reports the median tumor volume evaluation of H460 tumors inmice treated with placebo, ET-736 and 32. These results are also showedin FIG. 51.

TABLE 62 Median Tumor Volume (mm³) Days Control 32 ET-736 0 187.4 183.9185.8 2 577.5 455.2 457.8 5 1352.0 784.8 732.8 7 1642.9 837.4 930.1 92025.0 1044.3 1207.2 12 2025.0 1452.4 1568.0 14 1845.5 1845.5 16 2025.02025.0

Table 63 reports the median tumor volume evaluation of H526 tumors inmice treated with placebo, ET-736 and 32. These results are also showedin FIG. 52.

TABLE 63 Median Tumor Volume (mm³) Days Control 32 ET-736 0 217.2 212.1213.5 2 410.7 277.3 240.5 4 778.5 127.0 97.2 7 1083.2 95.0 48.8 9 1371.063.1 62.5 11 1782.0 62.5 62.5 14 2025.0 62.5 47.3 16 62.5 32.0 21 4.04.0 28 4.0 4.0 35 55.3 4.0 42 85.3 4.0 49 185.6 4.0 56 169.1 4.0 63 62.54.0 70 88.9 4.0 77 280.6 4.0 84 694.2 199.8 91 1150.9 786.5

Table 64 reports the median tumor volume evaluation of H82 tumors inmice treated with placebo, ET-736 and 32. These results are also showedin FIG. 53.

TABLE 64 Median Tumor Volume (mm³) Days Control 32 ET-736 0 171.6 171.6170.0 2 439.4 309.4 334.4 5 1024.7 485.0 539.4 7 1422.0 708.4 836.4 91923.8 972.6 1013.1 12 2025.0 1101.6 1290.9 14 1339.6 1648.0 16 1430.319 1885.7

Example 36d. In Vivo Studies to Determine the Effect of 32 in HumanOvarian Tumor Xenografts

The aim of this study was to compare the antitumoral activities of 32and ET-736 by using a xenograft model of human ovarian cancer.

The tumor model used in this study was A2780.

Table 65 reports the volume evaluation of A2780 tumors in mice treatedwith placebo, ET-736 and 32. These results are also showed in FIG. 54.

TABLE 65 Median Tumor Volume (mm³) Days Control 32 ET-736 0 169.5 168.6168.8 2 317.5 262.9 251.2 5 758.9 572.7 382.6 7 1351.9 997.5 676.1 91675.8 1359.9 959.4 12 2025.0 1715.0 1241.5 14 2025.0 1582.7 16 2025.01646.4 19 1845.5 21 2025.0

Example 36e. In Vivo Studies to Determine the Effect of 32 in HumanGastric Tumor Xenografts

The aim of this study was to compare the antitumoral activities of 32and ET-736 by using a xenograft model of human gastric cancer.

The tumor model used in this study was HGC27.

Table 66 reports tumor volume growth of HGC27 tumors in mice treatedwith placebo, ET-736 and 32. These results are also showed in FIG. 55.

TABLE 66 Median Tumor Volume (mm³) Days Control 32 ET-736 0 200.7 194.8195.9 2 429.0 386.3 359.2 5 835.5 551.3 537.6 7 1256.5 579.2 553.5 91602.2 665.8 604.7 12 2040.7 701.1 627.4 14 814.5 648.0 16 959.9 687.619 1312.4 760.0 21 1626.8 792.4 23 1737.3 818.9 26 1026.1 28 1354.9

Example 37. In Vivo Studies to Determine the Effect of 35 in SeveralXenograft Models

Compound 35 was provided in the form of freeze-dried vials oflyophilized product. Each vial was reconstituted with sterile water forinjection to a concentration of 0.5 mg/mL. Further dilutions were madewith 5% dextrose solution for injection to the dosing formulationconcentration. The administered dose of 35 was 0.25 mg/Kg.

PM01183 was provided in the form of vials of lyophilized product. Eachvial was reconstituted with sterile water for injection to aconcentration of 0.5 mg/mL. Further dilutions were made with 5% glucoseor 0.9% sodium chloride solution for injection to the dosing formulationconcentration. The administered dose of PM01183 was 0.18 mg/Kg.

Placebo was provided in the form of vials of lyophilised product eachvial (sucrose 200 mg+potassium dihydrogen phosphate 13.6 mg+phosphoricacid q.s. pH 3.8-4.5) was reconstituted with sterile water for injection(2 mL). Further dilutions were made with 5% dextrose solution forinjection.

In this experiment, compound 35 and PM01183, as well as placebo wereintravenously administered on a weekly schedule at a volume of 10 mL/Kg.

Example 37a. In Vivo Studies to Determine the Effect of 35 in HumanFibrosarcoma Xenografts

The aim of this study was to evaluate the antitumoral activities ofcompound 35 and PM01183 by using a xenograft model of human sarcoma.

The tumor model used in this study was HT-1080 cell line.

Table 67 reports the total diameter (tumor+leg) evaluation of HT1080tumors in mice treated with placebo, PM01183 and 35. These results arealso showed in FIG. 56.

TABLE 67 Total diameter (tumor + leg) (mm) Days Control PM01183 35 0 8.48.4 8.3 2 10.9 9.8 9.4 5 14.8 9.7 8.7 7 15.9 11.4 8.0 9 18.0 12.7 9.9 1213.7 11.4 14 14.6 12.5 16 15.5 13.2 19 15.6 14.6 21 18.0 15.7 23 18.0

Example 37b. In Vivo Studies to Determine the Effect of 35 in HumanBreast Adenocarcinoma Xenografts

The aim of this study was to compare the antitumoral activities of 35and PM01183 by using a xenograft model of human breast cancer.

The tumor model used in this study was MDA-MB-231 cell line.

Table 68 reports the median tumor volume evaluation of MDA-MB-231 tumorsin mice treated with placebo, PM01183 and 35. These results are alsoshowed in FIG. 57.

TABLE 68 Median Tumor Volume (mm³) Days Control 35 PM01183 0 132.6 132.7134.3 4 194.1 193.6 177.2 7 248.2 179.1 186.3 11 377.6 276.7 250.7 14461.3 286.0 266.1 18 679.2 384.5 327.7 21 753.2 436.8 391.0 25 909.2554.3 493.1 28 1090.7 647.0 627.3 32 1433.4 817.5 789.0 36 1887.5 1156.71022.0 39 1785.2 1387.6 1294.2 42 2081.5 1595.3 1643.3 46 2137.5 1689.91658.9 49 2044.2 1938.0

Example 37c. In Vivo Studies to Determine the Effect of 35 in Human LungCancer Xenografts

The aim of this study was to compare the antitumoral activities of 35and PM01183 by using a xenograft model of human lung cancer.

The tumor model used in this study was H460 cell line.

Table 69 reports the median tumor volume evaluation of H460 tumors inmice treated with placebo, PM01183 and 35. These results are also showedin FIG. 58.

TABLE 69 Median Tumor Volume (mm³) Days Control PM01183 35 0 197.0 196.3197.2 2 529.5 457.0 415.3 4 1057.4 861.5 750.8 7 1582.5 1280.2 1242.3 92094.8 1424.9 1536.3 11 1969.9 1728.7 14 2080.9

Example 37d. In Vivo Studies to Determine the Effect of 35 in HumanOvarian Tumor Xenografts

The aim of this study was to compare the antitumoral activities of 35and PM01183 by using a xenograft model of human ovarian cancer.

The tumor model used in this study was A2780.

Table 70 reports the volume evaluation of A2780 tumors in mice treatedwith placebo, PM01183 and 35. These results are also showed in FIG. 59.

TABLE 70 Median Tumor Volume (mm³) Days Control PM01183 35 0 163.4 163.6163.6 2 287.1 236.5 189.9 4 568.7 463.2 284.3 7 1211.3 986.3 606.4 91633.7 1451.4 946.9 11 2047.8 2062.0 1394.2 14 2067.7

Example 37e. In Vivo Studies to Determine the Effect of 35 in HumanGastric Tumor Xenografts

The aim of this study was to compare the antitumoral activities of 35and PM01183 by using a xenograft model of human gastric cancer.

The tumor model used in this study was HGC27.

Table 71 reports volume growth of HGC27 tumors in mice treated withplacebo, PM01183 and 35. These results are also showed in FIG. 60.

TABLE 71 Median Tumor Volume (mm³) Days Control 35 PM01183 0 178.3 182.3177.6 2 409.0 382.2 395.6 5 907.4 610.8 572.4 7 1283.6 775.5 766.6 91664.0 988.0 950.7 12 1692.4 1005.6 972.0 14 2102.8 1531.7 1199.4 161866.3 1353.1

Example 38. In Vivo Studies to Determine the Effect of 12-S and 12-R inHuman Prostate Xenografts

12-S and 12-R were provided in the form of freeze-dried vials oflyophilized product. Each vial was reconstituted with water for infusionto a concentration of 0.5 mg/mL. Further dilutions were made with 5%dextrose solution for injection to the dosing formulation concentration.The administered doses of 12-S and 12-R were 0.25 mg/kg and 0.05 mg/kgrespectively.

Placebo was provided in the form of lyophilised cake containing 100 mgSucrose+Potassium dihydrogen phosphate 6.8 mg+Phosphoric acid q.s. pH3.8-4.5 which was reconstituted with water for infusion.

In these experiments, 12-S and 12-R, as well as placebo, wereintravenously administered once per week for 3 consecutive weeks, onDays 0, 7 and 14, whenever it was possible.

The aim of this study was to compare the antitumoral activity of 12-Sand 12-R by using a xenograft model of human prostate cancer.

The tumor model used in this study was PC-3 cell line.

Table 72 reports the median tumor volume evaluation of PC-3 tumors inmice treated with placebo, 12-S and 12-R. These results are also showedin FIG. 61.

TABLE 72 Median Tumor Volume (mm³) Days Control 12-R 12-S 0 128.0 129.0128.0 2 149.6 136.2 141.5 4 197.0 144.2 143.7 7 250.9 172.2 183.9 11291.6 183.6 208.1 14 326.5 205.2 270.7 16 361.9 256.0 286.3 18 397.0325.7 336.1 21 476.9 322.2 357.1 23 506.1 407.8 400.8 25 526.7 419.9443.6 29 593.6 459.1 523.4 32 769.5 512.1 652.6 35 875.3 579.2 689.7 37900.0 613.8 692.2 39 977.8 764.1 726.9 42 1061.5 785.0 823.7 44 1463.4845.5 864.2 46 1612.8 748.0 1182.8 49 1809.2 808.7 1219.2 51 2030.9855.8 1331.9 56 1125.2 1335.2

Example 39. In Vivo Studies to Determine the Effect of 4-S in HumanProstate Xenografts

The aim of this study was to compare the antitumoral activity of 4-S byusing three different xenograft models of human prostate cancer. Thesemodels correspond to PC-3, DU-145 and 22Rv1 cell lines.

Compound 4-S was provided in the form of freeze-dried vials oflyophilized product. Each vial was reconstituted with sterile water forinjection to a concentration of 0.5 mg/mL. Further dilutions were madewith 5% dextrose solution for injection to the dosing formulationconcentration. The administered dose of 4-S varied depending on thestudy, being 1.25 mg/Kg when the tumor model was PC-3, 1.00 mg/Kg whenthe tumor model was DU-145 and 0.75 mg/Kg when the tumor model was22Rv1, respectively.

Placebo was provided in the form of lyophilised cake containing 100 mgSucrose+Potassium dihydrogen phosphate 6.8 mg+Phosphoric acid q.s. pH3.8-4.5 which was reconstituted with water for infusion.

In these experiments, 4-S, as well as placebo were intravenouslyadministered once per week for 3 consecutive weeks, on Days 0, 7 and 14,whenever it was possible.

Table 73 reports the median tumor volume evaluation of PC-3 tumors inmice treated with placebo and 4-S. These results are also showed in FIG.62.

TABLE 73 Median Tumor Volume (mm³) Days Control 4-S 0 140.5 141.3 2178.6 130.7 4 233.1 147.6 7 284.6 157.7 9 331.7 200.9 11 433.7 192.8 14500.4 210.8 16 570.8 255.5 18 680.3 261.1 21 850.1 282.4 23 928.5 382.225 915.7 451.6 28 1187.5 611.1 30 1270.1 762.3 32 1327.1 821.6 35 1373.61045.6

Table 74 reports the median tumor volume evaluation of DU-145 tumors inmice treated with placebo and 4-S. These results are also showed in FIG.63.

TABLE 74 Median Tumor Volume (mm³) Days Control 4-S 0 127.4 126.2 3180.9 102.4 5 248.8 119.5 7 320.4 149.5 10 384.6 216.8 12 441.0 181.4 14519.6 237.7 17 601.0 204.4 19 660.8 210.9 24 740.7 300.0 26 798.6 378.428 587.0 31 650.3

Table 75 reports the median tumor volume evaluation of 22Rv1 tumors inmice treated with placebo and 4-S. These results are also showed in FIG.64.

TABLE 75 Median Tumor Volume (mm³) Days Control 4-S 0 174.6 173.6 3307.2 70.3 5 511.5 63.1 7 739.1 76.7 10 955.2 49.1 12 1286.1 59.8 141385.8 74.9 17 1791.1 55.1 19 2025.0 64.9 24 138.4 26 186.9 28 242.0 31392.5 33 561.8 35 799.3 38 1107.0 40 1426.4 42 1685.5 45 2025.0

Example 40. In Vivo Studies to Determine the Effect of 39-S in HumanProstate Xenografts

The aim of this study was to compare the antitumoral activity of 39-S byusing three different xenograft models of human prostate cancer. Thesemodels correspond to PC-3, DU-145 and 22Rv1 cell lines.

Compound 39-S was provided in the form of freeze-dried vials oflyophilized product. Each vial was reconstituted with sterile water forinjection to a concentration of 0.5 mg/mL. Further dilutions were madewith 5% dextrose solution for injection to the dosing formulationconcentration. The administered dose of 39-S varied depending on thestudy, being 1.25 mg/Kg when the tumor model was PC-3, 1.00 mg/Kg whenthe tumor model was DU-145 and 0.75 mg/Kg when the tumor model was22Rv1, respectively.

Placebo was provided in the form of lyophilised cake containing 100 mgSucrose+Potassium dihydrogen phosphate 6.8 mg+Phosphoric acid q.s. pH3.8-4.5 which was reconstituted with water for infusion.

In these experiments, 39-S, as well as placebo, were intravenouslyadministered once per week for 3 consecutive weeks, on Days 0, 7 and 14,whenever it was possible.

Table 76 reports the median tumor volume evaluation of PC-3 tumors inmice treated with placebo and 39-S. These results are also showed inFIG. 65.

TABLE 76 Median Tumor Volume (mm³) Days Control 39-S 0 181.9 182.3 2254.8 222.6 4 308.7 244.0 7 344.5 269.3 9 396.8 295.8 11 439.2 315.0 14542.7 356.9 16 619.0 388.0 18 721.3 400.1 21 908.1 503.3 23 1039.1 556.025 1117.0 579.6 28 1232.3 694.9 30 1778.6 811.1 32 2018.1 1027.1 351194.3 37 1495.0 39 1710.7 42 2066.2

Table 77 reports the median tumor volume evaluation of DU-145 tumors inmice treated with placebo and 39-S. These results are also showed inFIG. 66.

TABLE 77 Median Tumor Volume (mm³) Days Control 39-S 0 156.8 179.9 2198.3 199.9 4 253.9 222.2 7 325.8 340.5 9 385.1 354.1 11 462.2 349.7 14483.8 429.1 16 599.0 454.8 18 664.0 449.7 21 816.9 517.5 23 861.3 568.525 977.9 629.4 28 973.6 775.7

Table 78 reports the median tumor volume evaluation of 22Rv1 tumors inmice treated with placebo and 39-S. These results are also showed inFIG. 67.

TABLE 78 Median Tumor Volume (mm³) Days Control 39-S 0 174.6 173.5 3307.2 93.0 5 511.5 96.8 7 739.1 115.2 10 955.2 108.2 12 1286.1 128.4 141385.8 155.6 17 1791.1 173.4 19 2025.0 210.2 24 358.8 26 456.5 28 645.231 1049.5 33 1439.4 35 2025.0

CLAUSES

1. A compound of formula I, or a pharmaceutically acceptable salt orester thereof:

wherein:

-   -   X is —NH— or —O—;    -   R₁ is —OH or —CN;    -   R₂ is a —C(═O)R^(a) group;    -   R₃ is hydrogen or a —OR^(b) group;    -   R₄ is selected from hydrogen, —CH₂OH, —CH₂O—(C═O)R^(c), —CH₂NH₂        and —CH₂NHProt^(NH);    -   R^(a) is selected from hydrogen, substituted or unsubstituted        C₁-C₁₂ alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl, and        substituted or unsubstituted C₂-C₁₂ alkynyl;    -   R^(b) is selected from substituted or unsubstituted C₁-C₁₂        alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl and        substituted or unsubstituted C₂-C₁₂ alkynyl;    -   R^(c) is selected from substituted or unsubstituted C₁-C₁₂        alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl, and        substituted or unsubstituted C₂-C₁₂ alkynyl; and    -   Prot^(NH) is a protecting group for amino;    -   with the proviso that when R₄ is hydrogen then X is —O—.        2. The compound according to clause 1 of formula IA or a        pharmaceutically acceptable salt or ester thereof:

wherein:

-   -   X is —NH— or —O—;    -   R₁ is —OH or —CN;    -   R₂ is a —C(═O)R^(a) group;    -   R₃ is hydrogen;    -   R₄ is selected from hydrogen, —CH₂OH, —CH₂O—(C═O)R^(c), —CH₂NH₂        and —CH₂NHProt^(NH);    -   R^(a) is selected from hydrogen, substituted or unsubstituted        C₁-C₁₂ alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl, and        substituted or unsubstituted C₂-C₁₂ alkynyl;    -   R^(c) is selected from substituted or unsubstituted C₁-C₁₂        alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl, and        substituted or unsubstituted C₂-C₁₂ alkynyl; and    -   Prot^(NH) is a protecting group for amino;    -   with the proviso that when R₄ is hydrogen then X is —O—.        3. The compound according to clause 1 of formula IB or a        pharmaceutically acceptable salt or ester thereof:

wherein:

-   -   X is —NH— or —O—;    -   R₁ is —OH or —CN;    -   R₂ is a —C(═O)R^(a) group;    -   R₃ is a —OR^(b) group;    -   R₄ is selected from hydrogen, —CH₂OH, —CH₂O—(C═O)R^(c), —CH₂NH₂        and —CH₂NHProt^(NH);    -   R^(a) is selected from hydrogen, substituted or unsubstituted        C₁-C₁₂ alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl, and        substituted or unsubstituted C₂-C₁₂ alkynyl;    -   R^(b) is selected from substituted or unsubstituted C₁-C₁₂        alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl and        substituted or unsubstituted C₂-C₁₂ alkynyl;    -   R^(c) is selected from substituted or unsubstituted C₁-C₁₂        alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl, and        substituted or unsubstituted C₂-C₁₂ alkynyl; and    -   Prot^(NH) is a protecting group for amino;    -   with the proviso that when R₄ is hydrogen then X is —O—.        4. The compound according to clause 1 selected from formula Ia        or Ib, or a pharmaceutically acceptable salt or ester thereof:

wherein:

-   -   X is —NH— or —O—;    -   R₁ is —OH or —CN;    -   R₂ is a —C(═O)R^(a) group;    -   R₃ is hydrogen or a —OR^(b) group;    -   R₄ is selected from —CH₂OH, —CH₂OC(═O)R^(c), —CH₂NH₂, and        —CH₂NHProt^(NH);    -   R^(a) is selected from hydrogen, substituted or unsubstituted        C₁-C₁₂ alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl, and        substituted or unsubstituted C₂-C₁₂ alkynyl;    -   R^(b) is selected from substituted or unsubstituted C₁-C₁₂        alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl, and        substituted or unsubstituted C₂-C₁₂ alkynyl;    -   R^(c) is selected from substituted or unsubstituted C₁-C₁₂        alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl, and        substituted or unsubstituted C₂-C₁₂ alkynyl; and    -   Prot^(NH) is a protecting group for amino.        5. The compound according to clause 2 selected from formula IAa        or IAb, or a pharmaceutically acceptable salt or ester thereof:

wherein:

-   -   X is —NH— or —O—;    -   R₁ is —OH or —CN;    -   R₂ is a —C(═O)R^(a) group;    -   R₃ is hydrogen;    -   R₄ is selected from —CH₂OH, —CH₂OC(═O)R^(c), —CH₂NH₂, and        —CH₂NHProt^(NH);    -   R^(a) is selected from hydrogen, substituted or unsubstituted        C₁-C₁₂ alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl, and        substituted or unsubstituted C₂-C₁₂ alkynyl;    -   R^(c) is selected from substituted or unsubstituted C₁-C₁₂        alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl, and        substituted or unsubstituted C₂-C₁₂ alkynyl; and    -   Prot^(NH) is a protecting group for amino.        6. The compound according to clause 3 selected from formula IBa        or IBb, or a pharmaceutically acceptable salt or ester thereof:

wherein:

X is —NH— or —O—; R₁ is —OH or —CN;

R₂ is a —C(═O)R^(a) group;R₃ is a —OR^(b) group;R₄ is selected from —CH₂OH, —CH₂OC(═O)R^(c), —CH₂NH₂, and—CH₂NHProt^(NH);R^(a) is selected from hydrogen, substituted or unsubstituted C₁-C₁₂alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl, and substituted orunsubstituted C₂-C₁₂ alkynyl;R^(b) is selected from substituted or unsubstituted C₁-C₁₂ alkyl,substituted or unsubstituted C₂-C₁₂ alkenyl, and substituted orunsubstituted C₂-C₁₂ alkynyl;R^(c) is selected from substituted or unsubstituted C₁-C₁₂ alkyl,substituted or unsubstituted C₂-C₁₂ alkenyl, and substituted orunsubstituted C₂-C₁₂ alkynyl; andProt^(NH) is a protecting group for amino.7. The compound according to any preceding clause wherein X is —NH—.8. The compound according to any preceding clause wherein X is —O—.9. The compound according to any preceding clause wherein R₄ is selectedfrom —CH₂OH, —CH₂O(C═O)R^(c), —CH₂NH₂, and —CH₂NHProt^(NH) wherein R^(c)is substituted or unsubstituted C₁-C₆ alkyl.10. The compound according to clause 9, wherein R^(c) is methyl.11. The compound according to clause 9, wherein R₄ is —CH₂OH.12. The compound according to clause 9, wherein R₄ is —CH₂NH₂.13. The compound according to clause 1 of formula Ic or apharmaceutically acceptable salt or ester thereof.

wherein:

R₁ is —OH or —CN;

R₂ is a —C(═O)R^(a) group;R₃ is hydrogen or a —OR^(b) group;R^(a) is selected from hydrogen, substituted or unsubstituted C₁-C₁₂alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl, and substituted orunsubstituted C₂-C₁₂ alkynyl; andR^(b) is selected from substituted or unsubstituted C₁-C₁₂ alkyl,substituted or unsubstituted C₂-C₁₂ alkenyl, and substituted orunsubstituted C₂-C₁₂ alkynyl.14. The compound according to clause 2 of formula IAc or apharmaceutically acceptable salt or ester thereof.

wherein:

R₁ is —OH or —CN;

R₂ is a —C(═O)R′ group;R₃ is hydrogen;R^(a) is selected from hydrogen, substituted or unsubstituted C₁-C₁₂alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl, and substituted orunsubstituted C₂-C₁₂ alkynyl.15. The compound according to clause 3 of formula IBc or apharmaceutically acceptable salt or ester thereof.

wherein:

R₁ is —OH or —CN;

R₂ is a —C(═O)R^(a) group;R₃ is a —OR^(b) group;R^(a) is selected from hydrogen, substituted or unsubstituted C₁-C₁₂alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl, and substituted orunsubstituted C₂-C₁₂ alkynyl; andR^(b) is selected from substituted or unsubstituted C₁-C₁₂ alkyl,substituted or unsubstituted C₂-C₁₂ alkenyl, and substituted orunsubstituted C₂-C₁₂ alkynyl.16. The compound according to any preceding clause wherein R₁ is —OH.17. The compound according to any preceding clause wherein R₂ is a—C(═O)R^(a) group where R^(a) is substituted or unsubstituted C₁-C₆alkyl.18. The compound according to clause 17 wherein R₂ is acetyl.19. The compound according to clause 1 wherein R₃ is hydrogen or —OR^(b)wherein R^(b) is substituted or unsubstituted C₁-C₆ alkyl.20. The compound according to clause 19 wherein R₃ is selected fromhydrogen and methoxy.21. The compound according to clause 20 wherein R₃ is hydrogen.22. The compound according to clause 20 wherein R₃ is methoxy.23. The compound according to any one of clauses 1, 2, 4, 5, 13 or 14wherein R₃ is hydrogen.24. The compound according to any one of clauses 1, 3, 4, 6, 13 or 15wherein R₃ is —OR^(b) wherein R^(b) is substituted or unsubstitutedC₁-C₆ alkyl.25. The compound according to clause 24 wherein R₃ is methoxy.26. The compound according to clause 1 of formula:

or a pharmaceutically acceptable salt or ester thereof.

27. The compound according to clause 1 of formula:

or a pharmaceutically acceptable salt or ester thereof.

28. The compound according to clause 1 of formula:

or a pharmaceutically acceptable salt or ester thereof.29. The compound according to clause 2 of formula:

or a pharmaceutically acceptable salt or ester thereof.

30. The compound according to clause 2 of formula:

a pharmaceutically acceptable salt or ester thereof.31. The compound according to clause 3 of formula:

or a pharmaceutically acceptable salt or ester thereof.

32. The compound according to clause 3 of formula:

or a pharmaceutically acceptable salt or ester thereof.

33. A pharmaceutical composition comprising a compound according to anyone of clauses 1 to 32 or a pharmaceutically acceptable salt or esterthereof and a pharmaceutically acceptable carrier.34. A compound according to any one of clauses 1 to 32, or apharmaceutically acceptable salt or ester thereof, or a compositionaccording to clause 33, for use as a medicament.35. A compound according to any one of clauses 1 to 32, or apharmaceutically acceptable salt or ester thereof, or a compositionaccording to clause 33, for use in the treatment of cancer.36. A method of treating cancer in a patient in need thereof, comprisingadministering to said patient a therapeutically effective amount ofcompound according to any one of clauses 1 to 32, or a pharmaceuticallyacceptable salt or ester thereof, or a composition according to clause33.37. The compound according to clause 35 or the method according toclause 36, wherein the cancer is selected from lung cancer includingnon-small cell lung cancer and small cell lung cancer, colon cancer,breast cancer, pancreas cancer, sarcoma, ovarian cancer, and gastriccancer.38. The compound or method according to clause 37, wherein the cancer isselected from lung cancer including non-small cell lung cancer and smallcell lung cancer, breast cancer, pancreas carcinoma and colorectalcancer.39. A process for obtaining a compound of formula I as defined in clause1 or a pharmaceutically acceptable salt or ester thereof, a compound offormula IA as defined in clause 2 or a pharmaceutically acceptable saltor ester thereof, or a compound of formula IB as defined in clause 3 ora pharmaceutically acceptable salt or ester thereof:comprising the step of reacting a compound of formula II with a compoundof formula III to give a compound of formula IV:

-   -   wherein:    -   X is —NH— or —O—;    -   R₂ is a —C(═O)R^(a) group;    -   R₃ is hydrogen or a —OR^(b) group;    -   R₄ is selected from hydrogen, —CH₂OH, —CH₂OC(═O)R^(c) and        —CH₂NHProt^(NH);    -   R^(a) is selected from hydrogen, substituted or unsubstituted        C₁-C₁₂ alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl,        substituted or unsubstituted C₂-C₁₂ alkynyl;    -   R^(b) is selected from substituted or unsubstituted C₁-C₁₂        alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl, and        substituted or unsubstituted C₂-C₁₂ alkynyl;    -   R^(c) is selected from substituted or unsubstituted C₁-C₁₂        alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl, and        substituted or unsubstituted C₂-C₁₂ alkynyl; and    -   Prot^(NH) is a protecting group for amino;    -   with the proviso that when R₄ is hydrogen then X is —O—.        40. The process according to clause 39, comprising the further        step of replacing the cyano group in the compound of formula IV        with a hydroxy group to give a compound of formula I, or IA or        IB where R₁ is OH.        41. A kit comprising a therapeutically effective amount of a        compound according to any one of clauses 1 to 32 and a        pharmaceutically acceptable carrier.        42. The kit according to clause 41 further comprising        instructions for use of the compound in the treatment of cancer,        and more preferably a cancer selected from lung cancer,        including non-small cell lung cancer and small cell lung cancer,        colon cancer, breast cancer, pancreas cancer, sarcoma, ovarian        cancer, and gastric cancer.        43. A compound of formula I, or a pharmaceutically acceptable        salt or ester thereof:

wherein:

-   -   X is —NH— or —O—;    -   R₁ is —OH or —CN;    -   R₂ is a —C(═O)R^(a) group;    -   R₃ is hydrogen or a —OR^(b) group;    -   R₄ is selected from hydrogen, —CH₂OH, —CH₂O—(C═O)R^(c), —CH₂NH₂        and —CH₂NHProt^(NH);    -   R^(a) is selected from hydrogen, substituted or unsubstituted        C₁-C₁₂ alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl, and        substituted or unsubstituted C₂-C₁₂ alkynyl;    -   R^(b) is selected from substituted or unsubstituted C₁-C₁₂        alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl and        substituted or unsubstituted C₂-C₁₂ alkynyl;    -   R^(c) is selected from substituted or unsubstituted C₁-C₁₂        alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl, and        substituted or unsubstituted C₂-C₁₂ alkynyl; and    -   Prot^(NH) is a protecting group for amino;    -   with the proviso that when R₄ is hydrogen then X is —O—.        44. The compound according to clause 43 selected from formula Ia        or Ib, or a pharmaceutically acceptable salt or ester thereof:

wherein:

X is —NH— or —O—; R₁ is —OH or —CN;

R₂ is a —C(═O)R^(a) group;R₃ is hydrogen or a —OR^(b) group;R₄ is selected from —CH₂OH, —CH₂OC(═O)R^(c), —CH₂NH₂, and—CH₂NHProt^(NH);R^(a) is selected from hydrogen, substituted or unsubstituted C₁-C₁₂alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl, and substituted orunsubstituted C₂-C₁₂ alkynyl;R^(b) is selected from substituted or unsubstituted C₁-C₁₂ alkyl,substituted or unsubstituted C₂-C₁₂ alkenyl, and substituted orunsubstituted C₂-C₁₂ alkynyl;R^(c) is selected from substituted or unsubstituted C₁-C₁₂ alkyl,substituted or unsubstituted C₂-C₁₂ alkenyl, and substituted orunsubstituted C₂-C₁₂ alkynyl; andProt^(NH) is a protecting group for amino.45. The compound according to clause 43 or clause 44 wherein X is —NH—.46. The compound according to clause 43 or clause 44 wherein X is —O—.47. The compound according to any one of clauses 43 to 46 wherein R₄ isselected from —CH₂OH, —CH₂O(C═O)R^(c), —CH₂NH₂, and —CH₂NHProt^(NH)wherein R^(c) is substituted or unsubstituted C₁-C₆ alkyl, preferablymethyl; particularly preferably wherein R₄ is —CH₂OH or —CH₂NH₂.48. The compound according to clause 43 of formula Ic or apharmaceutically acceptable salt or ester thereof

wherein:

R₁ is —OH or —CN;

R₂ is a —C(═O)R^(a) group;R₃ is hydrogen or a —OR^(b) group;R^(a) is selected from hydrogen, substituted or unsubstituted C₁-C₁₂alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl, and substituted orunsubstituted C₂-C₁₂ alkynyl; andR^(b) is selected from substituted or unsubstituted C₁-C₁₂ alkyl,substituted or unsubstituted C₂-C₁₂ alkenyl, and substituted orunsubstituted C₂-C₁₂ alkynyl.49. The compound according to any one of clauses 43 to 48 wherein R₁ is—OH; and/or wherein R₂ is a —C(═O)R^(a) group where R^(a) is substitutedor unsubstituted C₁-C₆ alkyl, preferably acetyl.50. The compound according to any one of clauses 43 to 49, wherein R₃ ishydrogen.51. The compound according to any one of clauses 43 to 49, wherein R₃ is—OR^(b); preferably wherein R^(b) is substituted or unsubstituted C₁-C₆alkyl, more preferably wherein R^(b) is methoxy.52. The compound according to clause 43 of formula:

or a pharmaceutically acceptable salt or ester thereof;

preferably of formula:

or a pharmaceutically acceptable salt or ester thereof.53. The compound according to clause 43 of formula:

or a pharmaceutically acceptable salt or ester thereof.54. The compound according to clause 45 of formula:

or a pharmaceutically acceptable salt or ester thereof;

preferably of formula:

or a pharmaceutically acceptable salt or ester thereof.55. A pharmaceutical composition comprising a compound according to anyone of clauses 43 to 53 or a pharmaceutically acceptable salt or esterthereof and a pharmaceutically acceptable carrier.56. A compound according to any one of clauses 43 to 54, or apharmaceutically acceptable salt or ester thereof, or a compositionaccording to clause 55, for use as a medicament; ora compound according to any one of clauses 43 to 54, or apharmaceutically acceptable salt or ester thereof, or a compositionaccording to clause 55, for use in the treatment of cancer; preferablywherein the cancer is selected from lung cancer including non-small celllung cancer and small cell lung cancer, colon cancer, breast cancer,pancreas cancer, sarcoma, ovarian cancer, and gastric cancer; even morepreferably wherein the cancer is selected from lung cancer includingnon-small cell lung cancer and small cell lung cancer, breast cancer,pancreas carcinoma and colorectal cancer.57. A process for obtaining a compound of formula I as defined in clause43 or a pharmaceutically acceptable salt or ester thereof:comprising the step of reacting a compound of formula II with a compoundof formula III to give a compound of formula IV:

-   -   wherein:    -   X is —NH— or —O—;    -   R₂ is a —C(═O)R^(a) group;    -   R₃ is hydrogen or a —OR^(b) group;    -   R₄ is selected from hydrogen, —CH₂OH, —CH₂OC(═O)R^(c) and        —CH₂NHProt^(NH);    -   R^(a) is selected from hydrogen, substituted or unsubstituted        C₁-C₁₂ alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl,        substituted or unsubstituted C₂-C₁₂ alkynyl;    -   R^(b) is selected from substituted or unsubstituted C₁-C₁₂        alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl, and        substituted or unsubstituted C₂-C₁₂ alkynyl;    -   R^(c) is selected from substituted or unsubstituted C₁-C₁₂        alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl, and        substituted or unsubstituted C₂-C₁₂ alkynyl; and    -   Prot^(NH) is a protecting group for amino;    -   with the proviso that when R₄ is hydrogen then X is —O—;

the process optionally comprising the further step of replacing thecyano group in the compound of formula IV with a hydroxy group to give acompound of formula I, or IA or IB where R₁ is OH.

1. A compound of formula I, or a pharmaceutically acceptable salt orester thereof:

wherein: X is —NH— or —O—; R₁ is —OH or —CN; R₂ is a —C(═O)R^(a) group;R₃ is hydrogen or a —OR^(b) group; R₄ is selected from hydrogen, —CH₂OH,—CH₂O—(C═O)R^(c), —CH₂NH₂ and —CH₂NHProt^(NH); R^(a) is selected fromhydrogen, substituted or unsubstituted C₁-C₁₂ alkyl, substituted orunsubstituted C₂-C₁₂ alkenyl, and substituted or unsubstituted C₂-C₁₂alkynyl; R^(b) is selected from substituted or unsubstituted C₁-C₁₂alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl and substituted orunsubstituted C₂-C₁₂ alkynyl; R^(c) is selected from substituted orunsubstituted C₁-C₁₂ alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl,and substituted or unsubstituted C₂-C₁₂ alkynyl; and Prot^(NH) is aprotecting group for amino; with the proviso that when R₄ is hydrogenthen X is —O—.
 2. The compound according to claim 1 selected fromformula Ia or Ib, or a pharmaceutically acceptable salt or esterthereof:

wherein: X is —NH— or —O—; R₁ is —OH or —CN; R₂ is a —C(═O)R^(a) group;R₃ is hydrogen or a —OR^(b) group; R₄ is selected from —CH₂OH,—CH₂OC(═O)R^(c), —CH₂NH₂, and —CH₂NHProt^(NH); R^(a) is selected fromhydrogen, substituted or unsubstituted C₁-C₁₂ alkyl, substituted orunsubstituted C₂-C₁₂ alkenyl, and substituted or unsubstituted C₂-C₁₂alkynyl; R^(b) is selected from substituted or unsubstituted C₁-C₁₂alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl, and substituted orunsubstituted C₂-C₁₂ alkynyl; R^(c) is selected from substituted orunsubstituted C₁-C₁₂ alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl,and substituted or unsubstituted C₂-C₁₂ alkynyl; and Prot^(NH) is aprotecting group for amino.
 3. The compound according to any one ofclaims 1 to 2, wherein X is —NH—.
 4. The compound according to any oneof claims 1 to 2, wherein X is —O—.
 5. The compound according to any oneof claims 1 to 4, wherein R₄ is selected from —CH₂OH, —CH₂O(C═O)R^(c),—CH₂NH₂, and —CH₂NHProt^(NH) wherein R^(c) is substituted orunsubstituted C₁-C₆ alkyl.
 6. The compound according to claim 5, whereinR^(c) is selected from substituted or unsubstituted methyl, substitutedor unsubstituted ethyl, substituted or unsubstituted n-propyl,substituted or unsubstituted isopropyl, substituted or unsubstitutedn-butyl, substituted or unsubstituted isobutyl, substituted orunsubstituted sec-butyl, and substituted or unsubstituted tert-butyl;preferably wherein R^(c) is methyl.
 7. The compound according to any oneof claim 1, or 3 when depended on claim 1, or 4 when depended on claim1, wherein R₄ is H, —CH₂OH or —CH₂NH₂.
 8. The compound according toclaim 5, wherein R₄ is —CH₂OH.
 9. The compound according to claim 5,wherein R₄ is —CH₂NH₂.
 10. The compound according to claim 1, of formulaIc or a pharmaceutically acceptable salt or ester thereof

wherein: R₁ is —OH or —CN; R₂ is a —C(═O)R^(a) group; R₃ is hydrogen ora —OR^(b) group; R^(a) is selected from hydrogen, substituted orunsubstituted C₁-C₁₂ alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl,and substituted or unsubstituted C₂-C₁₂ alkynyl; and R^(b) is selectedfrom substituted or unsubstituted C₁-C₁₂ alkyl, substituted orunsubstituted C₂-C₁₂ alkenyl, and substituted or unsubstituted C₂-C₁₂alkynyl.
 11. The compound according to any one of claims 1 to 10,wherein R₁ is —OH.
 12. The compound according to any one of claims 1 to11, wherein R₂ is a —C(═O)R^(a) group where R^(a) is substituted orunsubstituted C₁-C₆ alkyl; preferably wherein R^(a) is selected fromsubstituted or unsubstituted methyl, substituted or unsubstituted ethyl,substituted or unsubstituted n-propyl, substituted or unsubstitutedisopropyl, substituted or unsubstituted n-butyl, substituted orunsubstituted isobutyl, substituted or unsubstituted sec-butyl andsubstituted or unsubstituted tert-butyl.
 13. The compound according toclaim 12 wherein R₂ is acetyl.
 14. The compound according to any one ofclaims 1 to 13, wherein R₃ is hydrogen or —OR^(b) wherein R^(b) issubstituted or unsubstituted C₁-C₆ alkyl; preferably wherein R^(b) isselected from substituted or unsubstituted methyl, substituted orunsubstituted ethyl, substituted or unsubstituted n-propyl, substitutedor unsubstituted isopropyl, substituted or unsubstituted n-butyl,substituted or unsubstituted isobutyl, substituted or unsubstitutedsec-butyl and substituted or unsubstituted tert-butyl.
 15. The compoundaccording to claim 14 wherein R₃ is hydrogen.
 16. The compound accordingto claim 14 wherein R₃ is —OR^(b) wherein R^(b) is substituted orunsubstituted C₁-C₆ alkyl; preferably wherein R^(b) is selected fromsubstituted or unsubstituted methyl, substituted or unsubstituted ethyl,substituted or unsubstituted n-propyl, substituted or unsubstitutedisopropyl, substituted or unsubstituted n-butyl, substituted orunsubstituted isobutyl, substituted or unsubstituted sec-butyl andsubstituted or unsubstituted tert-butyl.
 17. The compound according toclaim 16 wherein R₃ is methoxy.
 18. The compound according to claim 1 offormula:

or a pharmaceutically acceptable salt or ester thereof.
 19. The compoundaccording to claim 1 of formula:

or a pharmaceutically acceptable salt or ester thereof.
 20. The compoundaccording to claim 1 of formula:

or a pharmaceutically acceptable salt or ester thereof.
 21. The compoundaccording to claim 1 of formula:

or a pharmaceutically acceptable salt or ester thereof.
 22. The compoundaccording to claim 1 of formula:

or a pharmaceutically acceptable salt or ester thereof.
 23. The compoundaccording to any one of claims 1 to 22, wherein the salt is selectedfrom hydrochloride, hydrobromide, hydroiodide, sulfate, nitrate,phosphate, acetate, trifluoroacetate, maleate, fumarate, citrate,oxalate, succinate, tartrate, malate, mandelate, methanesulfonate,p-toluenesulfonate, sodium, potassium, calcium, ammonium,ethylenediamine, ethanolamine, N,N-dialkylenethanolamine,triethanolamine and basic aminoacids.
 24. The compound according toclaim 1, of formula IC, or a pharmaceutically acceptable salt or esterthereof:

wherein: X is —NH—; R₁ is —OH or —CN; R₂ is a —C(═O)R^(a) group; R₃ ishydrogen or a —OR^(b) group; R₄ is selected from —CH₂OH,—CH₂O—(C═O)R^(c), —CH₂NH₂ and —CH₂NHProt^(NH); R^(a) is selected fromhydrogen, substituted or unsubstituted C₁-C₁₂ alkyl, substituted orunsubstituted C₂-C₁₂ alkenyl, and substituted or unsubstituted C₂-C₁₂alkynyl; R^(b) is selected from substituted or unsubstituted C₁-C₁₂alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl and substituted orunsubstituted C₂-C₁₂ alkynyl; R^(c) is selected from substituted orunsubstituted C₁-C₁₂ alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl,and substituted or unsubstituted C₂-C₁₂ alkynyl; and Prot^(NH) is aprotecting group for amino.
 25. The compound according to claim 24selected from formula ICa or ICb, or a pharmaceutically acceptable saltor ester thereof:

wherein: X is —NH—; R₁ is —OH or —CN; R₂ is a —C(═O)R^(a) group; R₃ ishydrogen or a —OR^(b) group; R₄ is selected from —CH₂OH,—CH₂OC(═O)R^(c), —CH₂NH₂, and —CH₂NHProt^(NH); R^(a) is selected fromhydrogen, substituted or unsubstituted C₁-C₁₂ alkyl, substituted orunsubstituted C₂-C₁₂ alkenyl, and substituted or unsubstituted C₂-C₁₂alkynyl; R^(b) is selected from substituted or unsubstituted C₁-C₁₂alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl, and substituted orunsubstituted C₂-C₁₂ alkynyl; R^(c) is selected from substituted orunsubstituted C₁-C₁₂ alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl,and substituted or unsubstituted C₂-C₁₂ alkynyl; and Prot^(NH) is aprotecting group for amino.
 26. The compound according to any one ofclaims 24 to 25, wherein R₄ is selected from —CH₂OH, —CH₂O(C═O)R^(c),—CH₂NH₂, and —CH₂NHProt^(NH) wherein R^(c) is substituted orunsubstituted C₁-C₆ alkyl.
 27. The compound according to claim 26,wherein is selected from substituted or unsubstituted methyl,substituted or unsubstituted ethyl, substituted or unsubstitutedn-propyl, substituted or unsubstituted isopropyl, substituted orunsubstituted n-butyl, substituted or unsubstituted isobutyl,substituted or unsubstituted sec-butyl, and substituted or unsubstitutedtert-butyl; preferably wherein R^(c) is methyl.
 28. The compoundaccording to claim 26, wherein R₄ is —CH₂OH or —CH₂NH₂.
 29. The compoundaccording to claim 26, wherein R₄ is —CH₂OH.
 30. The compound accordingto claim 26, wherein R₄ is —CH₂NH₂.
 31. The compound according to anyone of claims 24 to 30, wherein R₁ is —OH.
 32. The compound according toany one of claims 24 to 31, wherein R₂ is a —C(═O)R^(a) group whereR^(a) is substituted or unsubstituted C₁-C₆ alkyl; preferably whereinR^(a) is selected from substituted or unsubstituted methyl, substitutedor unsubstituted ethyl, substituted or unsubstituted n-propyl,substituted or unsubstituted isopropyl, substituted or unsubstitutedn-butyl, substituted or unsubstituted isobutyl, substituted orunsubstituted sec-butyl and substituted or unsubstituted tert-butyl. 33.The compound according to claim 32 wherein R₂ is acetyl.
 34. Thecompound according to any one of claims 24 to 33 wherein R₃ is hydrogenor —OR^(b) wherein R^(b) is substituted or unsubstituted C₁-C₆ alkyl;preferably wherein R^(b) is selected from substituted or unsubstitutedmethyl, substituted or unsubstituted ethyl, substituted or unsubstitutedn-propyl, substituted or unsubstituted isopropyl, substituted orunsubstituted n-butyl, substituted or unsubstituted isobutyl,substituted or unsubstituted sec-butyl and substituted or unsubstitutedtert-butyl.
 35. The compound according to claim 34 wherein R₃ ishydrogen.
 36. The compound according to claim 34 wherein R₃ is —OR^(b)wherein R^(b) is substituted or unsubstituted C₁-C₆ alkyl; preferablywherein R^(b) is selected from substituted or unsubstituted methyl,substituted or unsubstituted ethyl, substituted or unsubstitutedn-propyl, substituted or unsubstituted isopropyl, substituted orunsubstituted n-butyl, substituted or unsubstituted isobutyl,substituted or unsubstituted sec-butyl and substituted or unsubstitutedtert-butyl.
 37. The compound according to claim 36 wherein R₃ ismethoxy.
 38. The compound according to claim 24 of formula:

or a pharmaceutically acceptable salt or ester thereof.
 39. The compoundaccording to claim 24 of formula:

or a pharmaceutically acceptable salt or ester thereof.
 40. The compoundaccording to claim 24 of formula:

or a pharmaceutically acceptable salt or ester thereof.
 41. The compoundaccording to claim 24 of formula:

or a pharmaceutically acceptable salt or ester thereof.
 42. The compoundaccording to claim 24 of formula:

or a pharmaceutically acceptable salt or ester thereof.
 43. The compoundaccording to claim 1 of formula ID, or a pharmaceutically acceptablesalt or ester thereof:

wherein: X is —O—; R₁ is —OH or —CN; R₂ is a —C(═O)R^(a) group; R₃ ishydrogen or a —OR^(b) group; R₄ is selected from hydrogen, —CH₂OH,—CH₂O—(C═O)R^(c), —CH₂NH₂ and —CH₂NHProt^(NH); R^(a) is selected fromhydrogen, substituted or unsubstituted C₁-C₁₂ alkyl, substituted orunsubstituted C₂-C₁₂ alkenyl, and substituted or unsubstituted C₂-C₁₂alkynyl; R^(b) is selected from substituted or unsubstituted C₁-C₁₂alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl and substituted orunsubstituted C₂-C₁₂ alkynyl; R^(c) is selected from substituted orunsubstituted C₁-C₁₂ alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl,and substituted or unsubstituted C₂-C₁₂ alkynyl; and Prot^(NH) is aprotecting group for amino.
 44. The compound according to claim 43selected from formula IDa or IDb, or a pharmaceutically acceptable saltor ester thereof:

wherein: X is —O—; R₁ is —OH or —CN; R₂ is a —C(═O)R^(a) group; R₃ ishydrogen or a —OR^(b) group; R₄ is selected from —CH₂OH,—CH₂OC(═O)R^(c), —CH₂NH₂, and —CH₂NHProt^(NH); R^(a) is selected fromhydrogen, substituted or unsubstituted C₁-C₁₂ alkyl, substituted orunsubstituted C₂-C₁₂ alkenyl, and substituted or unsubstituted C₂-C₁₂alkynyl; R^(b) is selected from substituted or unsubstituted C₁-C₁₂alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl, and substituted orunsubstituted C₂-C₁₂ alkynyl; R^(c) is selected from substituted orunsubstituted C₁-C₁₂ alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl,and substituted or unsubstituted C₂-C₁₂ alkynyl; and Prot^(NH) is aprotecting group for amino.
 45. The compound according to any one ofclaims 43 to 44, wherein R₄ is selected from —CH₂OH, —CH₂O(C═O)R^(c),—CH₂NH₂, and —CH₂NHProt^(NH) wherein R^(c) is substituted orunsubstituted C₁-C₆ alkyl.
 46. The compound according to claim 45,wherein R^(c) is selected from substituted or unsubstituted methyl,substituted or unsubstituted ethyl, substituted or unsubstitutedn-propyl, substituted or unsubstituted isopropyl, substituted orunsubstituted n-butyl, substituted or unsubstituted isobutyl,substituted or unsubstituted sec-butyl, and substituted or unsubstitutedtert-butyl; preferably wherein R^(c) is methyl.
 47. The compoundaccording to claim 43, wherein R₄ is H, —CH₂OH or —CH₂NH₂.
 48. Thecompound according to claim 45, wherein R₄ is —CH₂OH.
 49. The compoundaccording to claim 45, wherein R₄ is —CH₂NH₂.
 50. The compound accordingto claim 43 of formula IDc or a pharmaceutically acceptable salt orester thereof

wherein: R₁ is —OH or —CN; R₂ is a —C(═O)R^(a) group; R₃ is hydrogen ora —OR^(b) group; R^(a) is selected from hydrogen, substituted orunsubstituted C₁-C₁₂ alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl,and substituted or unsubstituted C₂-C₁₂ alkynyl; and R^(b) is selectedfrom substituted or unsubstituted C₁-C₁₂ alkyl, substituted orunsubstituted C₂-C₁₂ alkenyl, and substituted or unsubstituted C₂-C₁₂alkynyl.
 51. The compound according to any one of claims 43 to 50,wherein R₁ is —OH.
 52. The compound according to any one of claims 43 to51, wherein R₂ is a —C(═O)R^(a) group where R^(a) is substituted orunsubstituted C₁-C₆ alkyl; preferably wherein R^(a) is selected fromsubstituted or unsubstituted methyl, substituted or unsubstituted ethyl,substituted or unsubstituted n-propyl, substituted or unsubstitutedisopropyl, substituted or unsubstituted n-butyl, substituted orunsubstituted isobutyl, substituted or unsubstituted sec-butyl andsubstituted or unsubstituted tert-butyl.
 53. The compound according toclaim 52 wherein R₂ is acetyl.
 54. The compound according to any one ofclaims 43 to 53, wherein R₃ is hydrogen or —OR^(b) wherein R^(b) issubstituted or unsubstituted C₁-C₆ alkyl; preferably wherein R^(b) isselected from substituted or unsubstituted methyl, substituted orunsubstituted ethyl, substituted or unsubstituted n-propyl, substitutedor unsubstituted isopropyl, substituted or unsubstituted n-butyl,substituted or unsubstituted isobutyl, substituted or unsubstitutedsec-butyl and substituted or unsubstituted tert-butyl.
 55. The compoundaccording to claim 54 wherein R₃ is hydrogen.
 56. The compound accordingto claim 54 wherein R₃ is —OR^(b) wherein R^(b) is substituted orunsubstituted C₁-C₆ alkyl; preferably wherein R^(b) is selected fromsubstituted or unsubstituted methyl, substituted or unsubstituted ethyl,substituted or unsubstituted n-propyl, substituted or unsubstitutedisopropyl, substituted or unsubstituted n-butyl, substituted orunsubstituted isobutyl, substituted or unsubstituted sec-butyl andsubstituted or unsubstituted tert-butyl.
 57. The compound according toclaim 56 wherein R₃ is methoxy.
 58. The compound according to claim 43of formula:

or a pharmaceutically acceptable salt or ester thereof.
 59. The compoundaccording to claim 43 of formula:

or a pharmaceutically acceptable salt or ester thereof.
 60. The compoundaccording to claim 43 of formula:

or a pharmaceutically acceptable salt or ester thereof.
 61. The compoundaccording to claim 43 of formula:

or a pharmaceutically acceptable salt or ester thereof.
 62. The compoundaccording to claim 1 of formula IE, or a pharmaceutically acceptablesalt or ester thereof:

wherein: X is —NH— or —O—; R₁ is —OH or —CN; R₂ is a —C(═O)R^(a) group;R₃ is hydrogen or a —OR^(b) group; R₄ is selected from —CH₂NH₂ and—CH₂NHProt^(NH); R^(a) is selected from hydrogen, substituted orunsubstituted C₁-C₁₂ alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl,and substituted or unsubstituted C₂-C₁₂ alkynyl; R^(b) is selected fromsubstituted or unsubstituted C₁-C₁₂ alkyl, substituted or unsubstitutedC₂-C₁₂ alkenyl and substituted or unsubstituted C₂-C₁₂ alkynyl; andProt^(NH) is a protecting group for amino.
 63. The compound according toclaim 62 selected from formula IEa or IEb, or a pharmaceuticallyacceptable salt or ester thereof:

wherein: X is —NH— or —O—; R₁ is —OH or —CN; R₂ is a —C(═O)R^(a) group;R₃ is hydrogen or a —OR^(b) group; R₄ is selected from —CH₂NH₂, and—CH₂NHProt^(NH); R^(a) is selected from hydrogen, substituted orunsubstituted C₁-C₁₂ alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl,and substituted or unsubstituted C₂-C₁₂ alkynyl; R^(b) is selected fromsubstituted or unsubstituted C₁-C₁₂ alkyl, substituted or unsubstitutedC₂-C₁₂ alkenyl, and substituted or unsubstituted C₂-C₁₂ alkynyl; andProt^(NH) is a protecting group for amino.
 64. The compound according toany one of claim 62 or 63 wherein X is —NH—.
 65. The compound accordingto any one of claim 62 or 63 wherein X is —O—.
 66. The compoundaccording to any one of claims 62 to 65, wherein R₄ is —CH₂NH₂.
 67. Thecompound according to any one of claims 62 to 66, wherein R₁ is —OH. 68.The compound according to any one of claims 62 to 67, wherein R₂ is a—C(═O)R^(a) group where R^(a) is substituted or unsubstituted C₁-C₆alkyl; preferably wherein R¹ is selected from substituted orunsubstituted methyl, substituted or unsubstituted ethyl, substituted orunsubstituted n-propyl, substituted or unsubstituted isopropyl,substituted or unsubstituted n-butyl, substituted or unsubstitutedisobutyl, substituted or unsubstituted sec-butyl and substituted orunsubstituted tert-butyl.
 69. The compound according to claim 68 whereinR₂ is acetyl.
 70. The compound according to any one of claims 62 to 69,wherein R₃ is hydrogen or —OR^(b) wherein R^(b) is substituted orunsubstituted C₁-C₆ alkyl; preferably wherein R^(b) is selected fromsubstituted or unsubstituted methyl, substituted or unsubstituted ethyl,substituted or unsubstituted n-propyl, substituted or unsubstitutedisopropyl, substituted or unsubstituted n-butyl, substituted orunsubstituted isobutyl, substituted or unsubstituted sec-butyl andsubstituted or unsubstituted tert-butyl.
 71. The compound according toclaim 70 wherein R₃ is hydrogen.
 72. The compound according to claim 70wherein R₃ is —OR^(b) wherein R^(b) is substituted or unsubstitutedC₁-C₆ alkyl; preferably wherein R^(b) is selected from substituted orunsubstituted methyl, substituted or unsubstituted ethyl, substituted orunsubstituted n-propyl, substituted or unsubstituted isopropyl,substituted or unsubstituted n-butyl, substituted or unsubstitutedisobutyl, substituted or unsubstituted sec-butyl and substituted orunsubstituted tert-butyl.
 73. The compound according to claim 72 whereinR₃ is methoxy.
 74. The compound according to claim 62 of formula:

or a pharmaceutically acceptable salt or ester thereof.
 75. The compoundaccording to claim 62 of formula:

or a pharmaceutically acceptable salt or ester thereof.
 76. The compoundaccording to claim 1 of formula IA or a pharmaceutically acceptable saltor ester thereof:

wherein: X is —NH— or —O—; R₁ is —OH or —CN; R₂ is a —C(═O)R^(a) group;R₃ is hydrogen; R₄ is selected from hydrogen, —CH₂OH, —CH₂O—(C═O)R^(c),—CH₂NH₂ and —CH₂NHProt^(NH); R^(a) is selected from hydrogen,substituted or unsubstituted C₁-C₁₂ alkyl, substituted or unsubstitutedC₂-C₁₂ alkenyl, and substituted or unsubstituted C₂-C₁₂ alkynyl; R^(c)is selected from substituted or unsubstituted C₁-C₁₂ alkyl, substitutedor unsubstituted C₂-C₁₂ alkenyl, and substituted or unsubstituted C₂-C₁₂alkynyl; and Prot^(NH) is a protecting group for amino; with the provisothat when R₄ is hydrogen then X is —O—.
 77. The compound according toclaim 76 selected from formula IAa or IAb, or a pharmaceuticallyacceptable salt or ester thereof:

wherein: X is —NH— or —O—; R₁ is —OH or —CN; R₂ is a —C(═O)R^(a) group;R₃ is hydrogen; R₄ is selected from —CH₂OH, —CH₂OC(═O)R^(c), —CH₂NH₂,and —CH₂NHProt^(NH); R^(a) is selected from hydrogen, substituted orunsubstituted C₁-C₁₂ alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl,and substituted or unsubstituted C₂-C₁₂ alkynyl; R^(c) is selected fromsubstituted or unsubstituted C₁-C₁₂ alkyl, substituted or unsubstitutedC₂-C₁₂ alkenyl, and substituted or unsubstituted C₂-C₁₂ alkynyl; andProt^(NH) is a protecting group for amino.
 78. The compound according toany one of claims 76 to 77, wherein X is —NH—.
 79. The compoundaccording to any one of claims 76 to 77, wherein X is —O—.
 80. Thecompound according to any one of claims 76 to 79, wherein R₄ is selectedfrom —CH₂OH, —CH₂O(C═O)R^(c), —CH₂NH₂, and —CH₂NHProt^(NH) wherein R¹ issubstituted or unsubstituted C₁-C₆ alkyl.
 81. The compound according toclaim 80, wherein R^(c) is selected from substituted or unsubstitutedmethyl, substituted or unsubstituted ethyl, substituted or unsubstitutedn-propyl, substituted or unsubstituted isopropyl, substituted orunsubstituted n-butyl, substituted or unsubstituted isobutyl,substituted or unsubstituted sec-butyl, and substituted or unsubstitutedtert-butyl; preferably wherein R¹ is methyl.
 82. The compound accordingto claim 76, or 78 when depended on claim 76, or 79 when depended onclaim 76, wherein R₄ is H, —CH₂OH or —CH₂NH₂.
 83. The compound accordingto claim 80, wherein R₄ is —CH₂OH.
 84. The compound according to claim80, wherein R₄ is —CH₂NH₂.
 85. The compound according to claim 76 offormula IAc or a pharmaceutically acceptable salt or ester thereof

wherein: R₁ is —OH or —CN; R₂ is a —C(═O)R^(a) group; R₃ is hydrogen;R^(a) is selected from hydrogen, substituted or unsubstituted C₁-C₁₂alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl, and substituted orunsubstituted C₂-C₁₂ alkynyl.
 86. The compound according to any one ofclaims 76 to 85, wherein R₁ is —OH.
 87. The compound according to anyone of claims 76 to 86, wherein R₂ is a —C(═O)R^(a) group where R^(a) issubstituted or unsubstituted C₁-C₆ alkyl; preferably wherein R^(a) isselected from substituted or unsubstituted methyl, substituted orunsubstituted ethyl, substituted or unsubstituted n-propyl, substitutedor unsubstituted isopropyl, substituted or unsubstituted n-butyl,substituted or unsubstituted isobutyl, substituted or unsubstitutedsec-butyl and substituted or unsubstituted tert-butyl.
 88. The compoundaccording to claim 87, wherein R₂ is acetyl.
 89. The compound accordingto claim 76 of formula:

or a pharmaceutically acceptable salt or ester thereof.
 90. The compoundaccording to claim 76 of formula:

or a pharmaceutically acceptable salt or ester thereof.
 91. The compoundaccording to claim 76 of formula:

or a pharmaceutically acceptable salt or ester thereof.
 92. The compoundaccording to claim 76 of formula:

or a pharmaceutically acceptable salt or ester thereof.
 93. The compoundaccording to claim 76 of formula:

or a pharmaceutically acceptable salt or ester thereof.
 94. The compoundaccording to claim 76 of formula:

or a pharmaceutically acceptable salt or ester thereof.
 95. The compoundaccording to claim 1 of formula IB or a pharmaceutically acceptable saltor ester thereof:

wherein: X is —NH— or —O—; R₁ is —OH or —CN; R₂ is a —C(═O)R^(a) group;R₃ is a —OR^(b) group; R₄ is selected from hydrogen, —CH₂OH,—CH₂O—(C═O)R^(c), —CH₂NH₂ and —CH₂NHProt^(NH); R^(a) is selected fromhydrogen, substituted or unsubstituted C₁-C₁₂ alkyl, substituted orunsubstituted C₂-C₁₂ alkenyl, and substituted or unsubstituted C₂-C₁₂alkynyl; R^(b) is selected from substituted or unsubstituted C₁-C₁₂alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl and substituted orunsubstituted C₂-C₁₂ alkynyl; R^(c) is selected from substituted orunsubstituted C₁-C₁₂ alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl,and substituted or unsubstituted C₂-C₁₂ alkynyl; and Prot^(NH) is aprotecting group for amino; with the proviso that when R₄ is hydrogenthen X is —O—.
 96. The compound according to claim 95 selected fromformula IBa or IBb, or a pharmaceutically acceptable salt or esterthereof:

wherein: X is —NH— or —O—; R₁ is —OH or —CN; R₂ is a —C(═O)R^(a) group;R₃ is a —OR^(b) group; R₄ is selected from —CH₂OH, —CH₂OC(═O)R^(c),—CH₂NH₂, and —CH₂NHProt^(NH); R^(a) is selected from hydrogen,substituted or unsubstituted C₁-C₁₂ alkyl, substituted or unsubstitutedC₂-C₁₂ alkenyl, and substituted or unsubstituted C₂-C₁₂ alkynyl; R^(b)is selected from substituted or unsubstituted C₁-C₁₂ alkyl, substitutedor unsubstituted C₂-C₁₂ alkenyl, and substituted or unsubstituted C₂-C₁₂alkynyl; R^(c) is selected from substituted or unsubstituted C₁-C₁₂alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl, and substituted orunsubstituted C₂-C₁₂ alkynyl; and Prot^(NH) is a protecting group foramino.
 97. The compound according to any one of claims 95 to 96, whereinX is —NH—.
 98. The compound according to any one of claims 95 to 96,wherein X is —O—.
 99. The compound according to any one of claims 95 to98, wherein R₄ is selected from —CH₂OH, —CH₂O(C═O)R^(c), —CH₂NH₂, and—CH₂NHProt^(NH) wherein R^(c) is substituted or unsubstituted C₁-C₆alkyl.
 100. The compound according to claim 99, wherein R¹ is selectedfrom substituted or unsubstituted methyl, substituted or unsubstitutedethyl, substituted or unsubstituted n-propyl, substituted orunsubstituted isopropyl, substituted or unsubstituted n-butyl,substituted or unsubstituted isobutyl, substituted or unsubstitutedsec-butyl, and substituted or unsubstituted tert-butyl; preferablywherein R^(c) is methyl.
 101. The compound according to claim 95, or 97when depended on claim 95, or 98 when depended on claim 95, wherein R₄is H, —CH₂OH or —CH₂NH₂.
 102. The compound according to claim 99,wherein R₄ is —CH₂OH.
 103. The compound according to claim 99, whereinR₄ is —CH₂NH₂.
 104. The compound according to claim 95, of formula IBcor a pharmaceutically acceptable salt or ester thereof

wherein: R₁ is —OH or —CN; R₂ is a —C(═O)R^(a) group; R₃ is a —OR^(b)group; R^(a) is selected from hydrogen, substituted or unsubstitutedC₁-C₁₂ alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl, andsubstituted or unsubstituted C₂-C₁₂ alkynyl; and R^(b) is selected fromsubstituted or unsubstituted C₁-C₁₂ alkyl, substituted or unsubstitutedC₂-C₁₂ alkenyl, and substituted or unsubstituted C₂-C₁₂ alkynyl. 105.The compound according to any one of claims 95 to 104, wherein R₁ is—OH.
 106. The compound according to any one of claims 95 to 105, whereinR₂ is a —C(═O)R^(a) group where R^(a) is substituted or unsubstitutedC₁-C₆ alkyl; preferably wherein R^(a) is selected from substituted orunsubstituted methyl, substituted or unsubstituted ethyl, substituted orunsubstituted n-propyl, substituted or unsubstituted isopropyl,substituted or unsubstituted n-butyl, substituted or unsubstitutedisobutyl, substituted or unsubstituted sec-butyl and substituted orunsubstituted tert-butyl.
 107. The compound according to claim 106wherein R₂ is acetyl.
 108. The compound according to any one of claims95 to 107, wherein R₃ is —OR^(b) wherein R^(b) is substituted orunsubstituted C₁-C₆ alkyl; preferably wherein R^(b) is selected fromsubstituted or unsubstituted methyl, substituted or unsubstituted ethyl,substituted or unsubstituted n-propyl, substituted or unsubstitutedisopropyl, substituted or unsubstituted n-butyl, substituted orunsubstituted isobutyl, substituted or unsubstituted sec-butyl andsubstituted or unsubstituted tert-butyl.
 109. The compound according toclaim 108 wherein R₃ is methoxy.
 110. The compound according to claim 95of formula:

or a pharmaceutically acceptable salt or ester thereof.
 111. Thecompound according to claim 95 of formula:

or a pharmaceutically acceptable salt or ester thereof.
 112. Thecompound according to claim 95 of formula:

or a pharmaceutically acceptable salt or ester thereof.
 113. A compoundaccording to any one of claims 24 to 112, wherein the salt is selectedfrom hydrochloride, hydrobromide, hydroiodide, sulfate, nitrate,phosphate, acetate, trifluoroacetate, maleate, fumarate, citrate,oxalate, succinate, tartrate, malate, mandelate, methanesulfonate,p-toluenesulfonate, sodium, potassium, calcium, ammonium,ethylenediamine, ethanolamine, N,N-dialkylenethanolamine,triethanolamine and basic aminoacids.
 114. A pharmaceutical compositioncomprising a compound according to any one of claims 1 to 113 or apharmaceutically acceptable salt or ester thereof and a pharmaceuticallyacceptable carrier.
 115. A dosage form comprising a pharmaceuticalcomposition according to claim
 114. 116. A compound according to any oneof claims 1 to 113, or a pharmaceutically acceptable salt or esterthereof, or a composition according to claim 114, or a dosage formaccording to claim 115, for use as a medicament.
 117. A compoundaccording to any one of claims 1 to 113, or a pharmaceuticallyacceptable salt or ester thereof, or a composition according to claim114, or a dosage form according to claim 115, for use in the treatmentof cancer.
 118. The compound, composition or dosage form according toclaim 117, wherein the cancer is selected from lung cancer includingnon-small cell lung cancer and small cell lung cancer, colon cancer,breast cancer, pancreas cancer, sarcoma, ovarian cancer, prostate cancerand gastric cancer.
 119. The compound, composition or dosage formaccording to claim 118, wherein the cancer is selected from lung cancerincluding non-small cell lung cancer and small cell lung cancer, breastcancer, pancreas cancer and colorectal cancer.
 120. Use of a compoundaccording to any one of claims 1 to 113, or a pharmaceuticallyacceptable salt or ester thereof, or a composition according to claim114, or a dosage form according to claim 115, in the manufacture of amedicament for the treatment of cancer.
 121. The use according to claim120, wherein the cancer is selected from lung cancer including non-smallcell lung cancer and small cell lung cancer, colon cancer, breastcancer, pancreas cancer, sarcoma, ovarian cancer, prostate cancer andgastric cancer.
 122. The use according to claim 121, wherein the canceris selected from lung cancer including non-small cell lung cancer andsmall cell lung cancer, breast cancer, pancreas cancer and colorectalcancer.
 123. A method of treating cancer in a patient in need thereof,comprising administering to said patient a therapeutically effectiveamount of compound according to any one of claims 1 to 113, or apharmaceutically acceptable salt or ester thereof, or a compositionaccording to claim 114, or a dosage form according to claim
 115. 124.The method according to claim 123, wherein the cancer is selected fromlung cancer including non-small cell lung cancer and small cell lungcancer, colon cancer, breast cancer, pancreas cancer, sarcoma, ovariancancer, prostate cancer and gastric cancer.
 125. The method according toclaim 124, wherein the cancer is selected from lung cancer includingnon-small cell lung cancer and small cell lung cancer, breast cancer,pancreas cancer and colorectal cancer.
 126. A process for obtaining acompound as defined in any one of claims 1 to 113 or a pharmaceuticallyacceptable salt or ester thereof: comprising the step of reacting acompound of formula II with a compound of formula III to give a compoundof formula IV:

wherein (where allowed by possible substituent groups): X is —NH— or—O—; R₂ is a —C(═O)R^(a) group; R₃ is hydrogen or a —OR^(b) group; R₄ isselected from hydrogen, —CH₂OH, —CH₂OC(═O)R^(c) and —CH₂NHProt^(NH);R^(a) is selected from hydrogen, substituted or unsubstituted C₁-C₁₂alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl, substituted orunsubstituted C₂-C₁₂ alkynyl; R^(b) is selected from substituted orunsubstituted C₁-C₁₂ alkyl, substituted or unsubstituted C₂-C₁₂ alkenyl,and substituted or unsubstituted C₂-C₁₂ alkynyl; R^(c) is selected fromsubstituted or unsubstituted C₁-C₁₂ alkyl, substituted or unsubstitutedC₂-C₁₂ alkenyl, and substituted or unsubstituted C₂-C₁₂ alkynyl; andProt^(NH) is a protecting group for amino; with the proviso that when R₄is hydrogen then X is —O—.
 127. The process according to claim 126,comprising the further step of replacing the cyano group in the compoundof formula IV with a hydroxy group to give a compound of formula I,where R₁ is OH.
 128. A kit comprising a therapeutically effective amountof a compound according to any one of claims 1 to 113 and apharmaceutically acceptable carrier.
 129. The kit according to claim 128further comprising instructions for use of the compound in the treatmentof cancer, and more preferably a cancer selected from lung cancer,including non-small cell lung cancer and small cell lung cancer, coloncancer, breast cancer, pancreas cancer, sarcoma, ovarian cancer,prostate cancer and gastric cancer.